Pyrazole derivatives and processes for the preparation thereof

ABSTRACT

4-Amino-1H-pyrazolo[3,4-d]pyrimidine derivatives of formula I ##STR1## wherein the symbols are as defined in claim 1, and intermediates for their manufacture are described. 
     The compounds of formula I inhibit especially the tyrosine kinase activity of the receptor for epidermal growth factor and can be used, for example, in the case of epidermal hyperproliferation (psoriasis) and as anti-tumor agents.

The invention relates to 4-amino-1H-pyrazolo[3,4-d]pyrimidinederivatives and intermediates and to processes for the preparationthereof, to pharmaceutical formulations comprising such derivatives, andto the use of those derivatives as medicaments.

The invention relates to 4-amino-1H-pyrazolo[3,4-d]pyrimidinederivatives of formula I ##STR2## wherein m and n are each independentlyof the other an integer from 0 up to and including 3,

v is 0 or 1,

R is hydrogen or lower alkyl,

R₁ is halogen, cyano, trifluoromethyl, hydroxy, lower alkoxy, loweralkanoyloxy, carboxy, lower alkoxycarbonyl, carbamoyl, N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by cyano, it being possible when several phenyl substituents R₁are present for those substituents to be identical or different from oneanother,

X is the group NH(CH--R₇)_(t) wherein t is an integer from 0 up to andincluding 3 and R₇ is hydrogen or lower alkyl, or the group (C[R₃]--R₄)_(q) wherein q is an integer from 0 up to and including 3, R₃ ishydrogen or lower alkyl and R₄ is hydrogen or lower alkyl, and R₂ ishalogen, nitro, cyano, trifluoromethyl, carboxy, lower alkoxycarbonyl,carbamoyl, N-lower alkylcarbamoyl, N,N-di-lower alkylcarbamoyl, azido,amino, lower alkylamino, di-lower alkylamino, di-lower alkylamino-loweralkyleneamino, benzylamino; acylated or sulfonated amino each having upto 10 carbon atoms; hydroxy, lower alkanoyloxy, oxa-lower alkoxy, loweralkoxy that is unsubstituted or substituted by carboxy, loweralkoxycarbonyl, carbamoyl or by N-lower alkylcarbamoyl, or lower alkylthat is unsubstituted or substituted by amino, lower alkanoylamino,benzoylamino, lower alkoxycarbonylamino, sulfonated amino, cyano,hydroxy, lower alkanoyloxy, lower alkoxycarbonyloxy or by lower alkoxy,it being possible when several phenyl substituents R₂ are present forthose substituents to be identical or different from one another and fortwo vicinal radicals R₂ together also to form methylenedioxy, and tosalts and tautomers of such compounds.

When m or n is 0, the phenyl ring in question does not carry asubstituent R₁ or R₂, respectively. Preferably, m and n are eachindependently of the other an integer from 0 up to and including 2. Whenm and/or n is/are 1, the phenyl substituent R₁ and/or R₂ is/areprimarily in the 4-position, i.e. in the para-position, or especially inthe 3-position, i.e. in the meta-position. When m and/or n is/are 2, thetwo phenyl substituents R₁ and/or R₂ are preferably in the 3- and4-positions.

When v is 0, the (R₁)_(m) phenyl radical is bonded directly to thenitrogen atom in the 4-position of the 1H-pyrazolo[3,4-d]pyrimidinederivative.

Halogen R₁ or R₂ is fluorine, bromine, iodine or, preferably, chlorine.

Lower alkoxy R₁ or R₂ is, for example, methoxy.

Lower alkanoyloxy R₁ or R₂ is, for example, acetoxy.

Lower alkoxycarbonyl R₁ or R2 is, for example, methoxycarbonyl.

N-Lower alkylcarbamoyl R₁ or R₂ is, for example, N-methylcarbamoyl.

Lower alkyl R₁ or R₂ that is substituted by amino or by cyano is, forexample, --(CH₂)_(x) --NH₂ or --(CH₂)_(x) --CN, wherein x is in eachcase from 1 to 4.

The group NH(CH--R₇)_(t) that is represented by the symbol X represents,when t is 0, the bivalent group NH. When t is from 1 to 3 and R₇ ishydrogen, the group NH(CH--R₇)_(t) represents the bivalent radicalsNHCH₂, NH--CH₂ --CH₂ and NH--CH₂ --CH₂ --CH₂, respectively, each ofwhich is bonded by its nitrogen atom to the pyrazole ring and by itsterminal carbon atom to the phenyl ring. When t is 1 and R₇ is loweralkyl, the group NH(CH--R₇)_(t) represents the bivalent radicalNH--CH(lower alkyl), for example the radical NH--CH(CH₃). X ispreferably NH, NH--CH₂ or NH--CH(CH₃).

The group (C[R₃ ]--R₄)_(q) that is represented by the symbol X is, whenq is 1, bonded by the underlined carbon atom (C[R₃ ]--R₄)_(q) both tothe pyrazole ring and to the phenyl ring and is preferably CH₂ orCH(lower alkyl). When q in the group (C[R₃ ]--R₄)_(q) is 2 or 3, the twofree valencies originate from different carbon atoms, such as, forexample, in dimethylene or trimethylene. When q is 0, the phenyl radicalis bonded directly to the pyrazole ring.

Lower alkylamino R₂ is, for example, methylamino.

Di-lower alkylamino-lower alkyleneamino R₂ is a radical of the formula

    --N═C(R.sub.8)--N(lower alkyl).sub.2,

wherein R₈ is hydrogen or lower alkyl, and is especially di-loweralkylaminomethyleneamino, such as dimethylaminomethyleneamino of theformula (CH₃)₂ N--CH═N--.

Acylated amino having up to 10 carbon atoms R₂ is, for example,unsubstituted or substituted lower alkanoylamino, benzoylamino, loweralkoxycarbonylamino, benzyloxycarbonylamino or monocyclic five- orsix-membered heterocyclylcarbonylamino. Unsubstituted loweralkanoylamino is, for example, formylamino, acetylamino, propionylamino,3-methylbutanoylamino or pivaloylamino. In such a substituted loweralkanoylamino radical the substituted lower alkanoyl radical is derivedpreferably from a naturally occurring amino acid, especially from one ofthe 20 amino acids that occur regularly in proteins, such as glycine,alanine, phenylalanine, etc. Preferred substituents in such asubstituted lower alkanoylamino radical R₂ are therefore amino and,possibly, also hydroxy, mercapto, methylthio, carboxy, carbamoyl,phenyl, 4-hydroxy-phenyl, imidazolyl or indolyl. Loweralkoxycarbonylamino is, for example, tert-butyloxycarbonylamino.Monocyclic five- or six-membered heterocyclylcarbonylamino is, forexample, thien-2-ylcarbonylamino, fur-2-ylcarbonylamino orpyrid-2-ylcarbonylamino.

Sulfonated amino having up to 10 carbon atoms R₂ is, for example, loweralkylsulfonylamino, such as especially methylsulfonylamino, orunsubstituted or substituted, for example lower alkyl-substituted,benzenesulfonylamino, such as p-toluenesulfonylamino.

Oxa-lower alkoxy is a lower alkoxy radical wherein one or more carbonatoms that are not adjacent to one another and that are other than C-1have been replaced by oxygen, for example --O--CH₂ --OCH₃, --O--CH₂--O--CH₂ --CH₃, --O--CH₂ --CH₂ --OCH₃ or --O--CH₂ --CH₂ --O--CH₂ --CH₂--OCH₃.

Lower alkoxy R₂ that is substituted by carboxy, lower alkoxycarbonyl,carbamoyl or by N-lower alkylcarbamoyl is, for example, carboxymethoxy,lower alkoxycarbonylmethoxy, carbamoylmethoxy, N-loweralkylcarbamoylmethoxy or ethoxy or n-propyloxy that is correspondinglysubstituted in the 2- or 3-position, respectively, i.e. in theω-position.

Lower alkyl R₂ that is substituted by amino or by hydroxy is, forexample, aminomethyl or hydroxymethyl.

Two vicinal radicals R₂ that together are methylenedioxy may be in the2,3- or 3,4-position. 2,3-Methylenedioxy-phenyl isbenzo[1,3]dioxol-4-yl. 3,4-Methylenedioxy-phenyl isbenzo-[1,3]dioxol-5-yl.

Within the context of the present invention, the general terms usedhereinbefore and hereinafter are preferably defined as follows:

The prefix "lower " denotes a radical having up to and including amaximum of 7, especially up to and including a maximum of 4, and aboveall 1 or 2, carbon atoms.

Halogen is preferably fluorine, chlorine, bromine or iodine, especiallyfluorine, chlorine or bromine, more especially bromine and above allchlorine.

Alkyl is unbranched or mono- or poly-branched and has preferably up to amaximum of 20 carbon atoms. Preference is given to lower alkyl,especially n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,tert-butyl, n-pentyl, neopentyl, n-hexyl, more especially ethyl andabove all methyl.

Alkoxy contains an alkyl radical as last defined and is especially loweralkoxy, such as n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy,tert-butoxy, n-pentyloxy, neopentyloxy, n-hexyloxy, especially ethoxyand above all methoxy.

Since compounds of formula I have basic properties, salts of thosecompounds are acid addition salts with organic or inorganic acids,especially the pharmaceutically acceptable, non-toxic salts. Suitableinorganic acids are, for example, carbonic acid (preferably in the formof the carbonates or hydrogen carbonates); hydrohalic acids, such ashydrochloric acid; sulfuric acid; or phosphoric acid. Suitable organicacids are, for example, carboxylic, phosphonic, sulfonic or sulfamicacids, for example acetic acid, propionic acid, octanoic acid, decanoicacid, dodecanoic acid, glycolic acid, lactic acid, 2-hydroxybutyricacid, gluconic acid, glucosemonocarboxylic acid, fumaric acid, succinicacid, adipic acid, pimelic acid, suberic acid, azelaic acid, malic acid,tartaric acid, citric acid, glucaric acid, galactaric acid, amino acids,such as glutamic acid, aspartic acid, N-methylglycine, acetylaminoaceticacid, N-acetylasparagine or N-acetylcystine, pyruvic acid, acetoaceticacid, phosphoserine, 2- or 3-glycerophosphoric acid,glucose-6-phosphoric acid, glucose-1-phosphoric acid,fructose-1,6-bis-phosphoric acid, maleic acid, hydroxymaleic acid,methylmaleic acid, cyclohexanecarboxylic acid, adamantanecarboxylicacid, benzoic acid, salicylic acid, 1- or 3-hydroxynaphthyl-2-carboxylicacid, 3,4,5-trimethoxybenzoic acid, 2-phenoxybenzoic acid,2-acetoxybenzoic acid, 4-aminosalicylic acid, phthalic acid,phenylacetic acid, mandelic acid, cinnamic acid, nicotinic acid,isonicotinic acid, glucuronic acid, galacturonic acid, methane orethane-sulfonic acid, 2-hydroxyethanesulfonic acid,ethane-1,2-disulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonicacid, 1,5-naphthalene-disulfonic acid, 2-, 3- or 4-methylbenzenesulfonicacid, methylsulfuric acid, ethylsulfuric acid, dodecylsulfuric acid,N-cyclohexylsulfamic acid, N-methyl-, N-ethyl- or N-propyl-sulfamicacid, or other organic protonic acids, such as ascorbic acid.

For isolation or purification purposes it is also possible to usepharmaceutically unacceptable salts, for example picrates orperchlorates. Only salts that are pharmaceutically acceptable andnon-toxic (at the appropriate dosages) are used therapeutically andthose salts are therefore preferred.

It is possible that the compounds of the formula I and the intermediatescontaining the pyrazole moiety are present under certain conditions,e.g. when dissolved in certain solvents, to some extent in a tautomericform wherein the nitrogen normally located at the nitrogen atom inposition 1 of the pyrazole moiety is translocated to another suitablenitrogen atom, e.g. the nitrogen atoms in positions 2, 5 or 7 of formulaI. The invention relates also to these tautomers.

The compounds of formula I have valuable pharmacologically usefulproperties. In particular they exhibit specific inhibitory activitiesthat are of pharmacological interest. They are effective especially asprotein tyrosine kinase inhibitors and/or (furthermore) as inhibitors ofprotein serine/threonine kinases; they exhibit, for example, powerfulinhibition of the tyrosine kinase activity of the receptor for epidermalgrowth factor (EGF) and of c-erbB2-kinase. Those receptor-specificenzyme activities play a key role in signal transmission in a largenumber of mammalian cells, including human cells, especially epithelialcells, cells of the immune system and cells of the central andperipheral nervous system. For example, in various cell types,EGF-induced activation of receptor-associated protein tyrosine kinase(EGF-R-PTK) is a prerequisite for cell division and hence for theproliferation of the cell population. The administration ofEGF-receptor-specific tyrosine kinase inhibitors thus inhibits theproliferation of the cells. The same applies analogously to the otherprotein kinases mentioned hereinbefore and hereinafter.

The inhibition of EGF-receptor-specific protein tyrosine kinase(EGF-R-PTK) can be demonstrated using known methods, for example usingthe recombinant intracellular domain of the EGF-receptor (EGF-R ICD;see, for example, E. McGlynn et al., Europ. J. Biochem. 207, 265-275(1992)). Compared with the control without inhibitor, the compounds offormula I inhibit the enzyme activity by 50% (IC₅₀), for example in aconcentration of from 0.0005 to 5 μM, especially from 0.001 to 0.1 μM.

In addition to or instead of inhibiting EGF-receptor tyrosine proteinkinase, the compounds of formula I also inhibit to a varying extentother tyrosine protein kinases that are involved in signal transmissionmediated by trophic factors, for example abl kinase, especially v-ablkinase, kinases from the family of the src kinases, especially c-srckinase, lck, fyn; further kinases of the EGF family, for example c-erbB2kinase (HER-2), c-erbB3 kinase, c-erbB4 kinase; members of the PDGFreceptor tyrosine protein kinase family, for example the PDGF receptorkinase, the (CSF-1 receptor kinase, the Kit receptor kinase, the VEGFreceptor kinase and the FGF receptor kinase; the insulin-like growthfactor receptor kinase (IGF-1 kinase), as well as serine/threoninekinases, for example protein kinase C or CDC kinases, all of which playa part in growth regulation and transformation in mammalian cells,including human cells.

The inhibition of c-erbB2-tyrosine kinase (HER-2) can be determined, forexample, analogously to the method used for EGF-R-PTK (see C. House etal., Europ. J. Biochem. 140, 363-367 (1984)). The c-erbB2-kinase can beisolated, and its activity determined, by means of protocols known perse, for example in accordance with T. Akiyama et al., Science 232, 1644(1986).

In the micromolar range too, the compounds of formula I exhibit, forexample, inhibition of the cell growth of EGF-dependent cell lines, forexample the epidermoid BALB/c mouse keratinocyte cell line (seeWeissmann, B. A., and Aaronson, S. A., Cell 32, 599 (1983)) or the A431cell line, which are recognized useful standard sources of EGF-dependentepithelial cells (see Carpenter, G., and Zendegni, J. Anal. Biochem.153, 279-282 (1985)). In a known test method (see Meyer et al., Int. J.Cancer 43, 851 (1989)), the inhibitory activity of the compounds offormula I is determined, briefly, as follows: BALB/MK cells (10000/microtiter plate well) are transferred to 96-well microtiter plates.The test compounds (dissolved in DMSO) are added in a series ofconcentrations (dilution series) in such a manner that the finalconcentration of DMSO is not greater than 1% (v/v). After the addition,the plates are incubated for three days during which the controlcultures without test compound are able to undergo at least threecell-division cycles. The growth of the MK cells is measured by means ofmethylene blue staining: after the incubation the cells are fixed withglutaraldehyde, washed with water and stained with 0.05% methylene blue.After a washing step the stain is eluted with 3% HCl and the opticaldensity per well of the microtiter plate is measured using a Titertekmultiskan at 665 nm. IC₅₀ values are determined by a computer-aidedsystem using the formula:

    IC.sub.50 =[(OD.sub.test -OD.sub.start)/(OD.sub.control -OD.sub.start)]×100.

The IC₅₀ value in those experiments is given as that concentration ofthe test compound in question that results in a cell count that is 50%lower than that obtained using the control without inhibitor. Thecompounds of formula I exhibit inhibitory activity in the micromolarrange, for example an IC₅₀ of approximately from 0.1 to 10 μM,especially from 0.4 to 4 μM.

The compounds of formula I exhibit inhibition of the growth of tumorcells also in vivo, as shown, for example, by the test described below:the test is based on inhibition of the growth of the human epidermoidcarcinoma A431 (ATCC No. CRL 1555; American Type Culture Collection,Rockville, Md., USA; see Santon, J. B., et al., Cancer Research 46,4701-4705 (1986) and Ozawa, S., et al., Int. J. Cancer 40, 706-710(1987)), which is transplanted into female BALB/c nude mice(Bomholtgard, Denmark). That carcinoma exhibits a growth that correlateswith the extent of the expression of EGF-receptor. In the experiment,tumors having a volume of approximately 1 cm³ cultured in vivo aresurgically removed from experimental animals under sterile conditions.The tumors are comminuted and suspended in 10 volumes (w/v) ofphosphate-buffered saline. The suspension is injected s.c. (0.2 ml/mousein phosphate-buffered saline) into the left flank of the animals.Alternatively, 1×10⁶ cells from an in vitro culture can be injected in0.2 ml of phosphate-buffered saline. Treatment with test compounds offormula I is started 5 or 7 days after the transplant, when the tumorshave reached a diameter of 4-5 mm. The test compound in question isadministered (in different doses for different animal groups) once a dayfor 15 successive days. The tumor growth is determined by measuring thediameter of the tumors along three axes that are perpendicular to eachother. The tumor volumes are calculated using the known formula p×L×D²/6 (see Evans, B. D., et al., Brit. J. Cancer 45, 466-8 (1982)). Theresults are given as treatment/control percentages (T/C×100=T/C %). At adose of from 3 to 50 mg/kg of active ingredient, distinct inhibition ofthe tumor growth is found, for example T/C % values of less than 10,which indicates strong inhibition of tumor growth.

The compounds of formula I which inhibit the tyrosine kinase activity ofthe receptor for the epidermal growth factor (EGF) or also of the otherprotein kinases mentioned are therefore useful, for example, in thetreatment of benign or malignant tumors. They are capable of effectingtumor regression and of preventing the formation of tumor metastases,the growth of micrometastases and the formation of new blood vessels(angiogenesis) required for tumor growth. They can be used especially inthe case of epidermal hyperproliferation (psoriasis), in the treatmentof neoplasia of epithelial character, e.g. mammary or ovariancarcinomas, and in leukemias. In addition, the compounds of formula I(especially the novel compounds) can be used in the treatment ofdisorders of the immune system in which several or, preferably,individual protein tyrosine kinases and/or (furthermore) proteinserine/threonine kinases are involved; those compounds of formula I canalso be used in the treatment of disorders of the central or peripheralnervous system in which signal transmission by several or, preferably, asingle protein tyrosine kinase(s) and/or (furthermore) proteinserine/threonine kinase(s) is/are involved.

In general, the present invention relates also to the use of thecompounds of formula I in the inhibition of the mentioned proteinkinases.

The compounds according to the invention can be used both alone and incombination with other pharmacologically active compounds, for exampletogether with inhibitors of the enzymes of polyamine synthesis,inhibitors of protein kinase C, inhibitors of other tyrosine kinases,cytokines, negative growth regulators, for example TGF-β or IFN-β,aromatase inhibitors, antiestrogens and/or cytostatic agents.

In the case of the preferred subjects of the invention mentionedhereinafter, general definitions can be replaced by the more specificdefinitions given at the beginning, where appropriate and expedient.

Preference is given to compounds of formula I according to claim 1,wherein R₂ is halogen, nitro, cyano, trifluoromethyl, carboxy, loweralkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, N,N-di-loweralkylcarbamoyl, azido, amino, lower alkylamino, di-lower alkylamino,di-lower alkylamino-lower alkyleneamino, benzylamino, benzoylamino,lower alkanoylamino, lower alkoxycarbonylamino, benzyloxycarbonylamino,thien-2-ylcarbonylamino, fur-2-ylcarbonylamino, pyrid-2-ylcarbonylamino,lower alkylsulfonylamino, benzenesulfonylamino, p-toluenesulfonylamino,hydroxy, lower alkanoyloxy, oxa-lower alkoxy, lower alkoxy that isunsubstituted or substituted by carboxy, lower alkoxycarbonyl, carbamoylor by N-lower alkylcarbamoyl, or lower alkyl that is unsubstituted orsubstituted by amino, lower alkanoylamino, benzoylamino, loweralkoxycarbonylamino, lower alkylsulfonylamino, benzenesulfonylamino,p-toluenesulfonylamino, cyano, hydroxy, lower alkanoyloxy, loweralkoxycarbonyloxy or by lower alkoxy, it being possible when severalphenyl substituents R₂ are present for those substituents to beidentical or different from one another and for two vicinal radicals R₂together also to form methylenedioxy, and the remaining symbols are asdefined above, and to salts of such compounds.

Preference is given to compounds of formula I according to claim 1,wherein

m is 0 or 1,

n is an integer from 0 up to and including 3,

v is 0or 1,

R is hydrogen or lower alkyl,

R₁ is halogen or lower alkyl that is unsubstituted or substituted byamino or by cyano,

X is the group NH(CH--R₇)_(t) wherein t is 0 or 1 and R₇ is hydrogen orlower alkyl, or the group (C[R₃ ]--R₄)_(q) wherein q is 0, and

R₂ is halogen, nitro, carboxy, lower alkoxycarbonyl, carbamoyl, N-loweralkylcarbamoyl, N,N-di-lower alkylcarbamoyl, amino, lower alkyiamino,di-lower alkylamino, di-lower alkylaminomethyleneamino, benzylamino,benzoylamino, lower alkanoylamino, lower alkoxycarbonylamino,benzyloxycarbonylamino, thien-2-yl-carbonylamino, fur-2-ylcarbonylamino,pyrid-2-ylcarbonylamino, lower alkylsulfonylamino, benzenesulfonylamino,p-toluenesulfonylamino, hydroxy, lower alkanoyloxy, lower alkoxy, orlower alkyl that is unsubstituted or substituted by amino, loweralkanoylamino, benzoylamino or by lower alkoxycarbonylamino, it beingpossible when several phenyl substituents R₂ are present for thosesubstituents to be identical or different from one another and for twovicinal radicals R₂ together also to form methylenedioxy,

and to salts of such compounds.

Preference is given also to 4-phenylamino-1H-pyrazolo[3,4-d]pyrimidinederivatives of formula Ia ##STR3## wherein m and n are eachindependently of the other an integer from 0 up to and including 3,

R₁ is halogen, cyano, trifluoromethyl, hydroxy, lower alkoxy, loweralkanoyloxy, carboxy, lower alkoxycarbonyl, carbamoyl, N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by cyano, it being possible when several phenyl substituents R₁are present for those substituents to be identical or different from oneanother,

X is the group NH(CH₂)_(t) wherein t is an integer from 0 up to andincluding 3, or the group (C[R₃ ]--R₄)_(q) wherein q is an integer from0 up to and including 3, R₃ is hydrogen or lower alkyl and R₄ ishydrogen or lower alkyl, and

R₂ is halogen, nitro, cyano, trifluoromethyl, carboxy, loweralkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, amino, loweralkylamino, acylated amino having up to 10 carbon atoms, hydroxy, loweralkanoyloxy, oxa-lower alkoxy, lower alkoxy that is unsubstituted orsubstituted by carboxy, lower alkoxycarbonyl, carbamoyl or by N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by hydroxy, it being possible when several phenyl substituentsR₂ are present for those substituents to be identical or different fromone another,

and to salts of such compounds.

Preference is given to compounds of formula Ia wherein X is the groupNH(CH₂)_(t) wherein t is an integer from 0 up to and including 3, or thegroup (C[R₃ ]--R₄)_(q) wherein q is 0, and the remaining symbols are asdefined above, and to salts of those compounds.

Preference is given especially to compounds of formula Ia wherein X isthe group NH(CH₂)_(t) wherein t is 0, and the remaining symbols are asdefined above, and to salts of those compounds.

Special preference is given to compounds of formula Ia wherein m and nare each independently of the other 0 or 1, R₁ is halogen, or loweralkyl that is unsubstituted or substituted by amino or by cyano, X isthe group NH(CH₂)_(t) wherein t is 0, and R₂ is halogen or lower alkoxy,and to salts of those compounds.

Special preference is given also to compounds of formula Ia wherein m is1 and R₁ is 3-chloro, and to salts of those compounds.

Preference is given above all to the compounds of formula I mentioned inthe Examples and to the pharmaceutically acceptable salts thereof.

The compounds of formula I and the salts thereof can be prepared in amanner known per se. The preparation process according to the inventioncomprises

a) treating a compound of formula II ##STR4## wherein R₅ is hydrogen ormethyl, R₆ is alkoxy having from 1 to 3 carbon atoms or is nitro, r isan integer from 0 to 2, and the remaining substituents and symbols areas defined above, with a suitable Lewis acid, or

b) reacting a compound of formula XV ##STR5## wherein the symbols are asdefined above, with an amine of formula XVI ##STR6## wherein v is 1 andthe remaining symbols are as defined above, or with a salt thereof, inthe presence of formic acid, or

c) reacting a compound of formula XV ##STR7## wherein the symbols are asdefined above, with a formamide derivative of formula XVII ##STR8##wherein v is 1 and the remaining symbols are as defined above, or d)reacting a compound of formula XVIII ##STR9## wherein the symbols are asdefined above, with an amine of formula XVI ##STR10## wherein v is 0 or1 and the remaining symbols are as defined above, or with a saltthereof, or

e) subjecting a compound of formula XIX ##STR11## wherein the symbolsare as defined above, to the conditions of a Dimroth rearrangement, and,if desired, converting a compound of formula I obtainable in accordancewith any one of processes a) to e) into its salt, or converting anobtainable salt of a compound of formula I into the free compound.

The procedure for those process variants and the preparation of thestarting materials are described in more detail below:

GENERAL POINTS

If necessary, interfering functional groups in starting materials areprotected before the reaction, in a manner known per se, by readilyremovable protecting groups which are removed again when the reactionhas taken place.

Process a):

When R₅ is hydrogen, a suitable Lewis acid is especially aluminumchloride. The reaction is carried out in an inert organic solvent, forexample a hydrocarbon, such as, preferably, an aromatic hydrocarbon,such as especially benzene or toluene, at a temperature of from roomtemperature (approx. 20° C.) to +200° C., if necessary under aprotective gas, such as argon, and/or under increased pressure,preferably at the boiling temperature of the solvent used, that is tosay under reflux. When R₅ is methyl, boiling is preferably carried outwith polyphosphoric acid.

The starting material of formula II is obtained as follows: first acompound of formula III ##STR12## wherein X, R₂ and n are as definedabove, is reacted with a hydrazine derivative of formula IV ##STR13##wherein R₅ is hydrogen or methyl, R₆ is alkoxy having from 1 to 3 carbonatoms or is nitro, and r is an integer from 0 to 2, or (in the presenceof a suitable base, like sodium methylate) with a salt thereof, to forma pyrazole derivative of formula V ##STR14## wherein the substituentsare as defined above. For example, there is used as starting material asolution in methanol of a hydrazine derivative of formula IV in the formof the dihydrochloride, to which there is added first, with cooling, forexample with ice, a solution of sodium methanolate in methanol and then,at room temperature, a solution of a compound of formula III in asuitable anhydrous alcohol, such as absolute ethanol. Heating is thencarried out for several hours under reflux.

The resulting compound of formula V is reacted with formic acid withsynthesis of the pyrimidine ring to form a compound of formula VI##STR15## wherein the substituents are as defined above. Preferably, acompound of formula V is heated under reflux for several hours in 85%aqueous formic acid.

From a compound of formula VI there is obtained with phosphoryl chloride(phosphorus oxychloride, POCl₃) or phosphorus trichloride (PCl₃), withreplacement of the hydroxy group by chlorine, a compound of formula VII##STR16## wherein the substituents are as defined above. Preferably, acompound of formula VI is heated under reflux for several hours inphosphoryl chloride under a protective gas, such as argon.

A compound of formula VII is then reacted with an aniline derivative offormula VIII ##STR17## wherein the symbols are as defined above,preferably in a suitable solvent, such as a suitable alcohol, e.g.ethanol, under a protective gas, such as nitrogen, at elevatedtemperature, for example under reflux, to form the desired startingmaterial of formula II.

The starting material of formula III wherein X is the group NH(CH₂)_(t)wherein t is as defined above is obtained, for example, by reacting3,3-bis-methylmercapto-2-cyano-acrylonitrile of formula IX ##STR18##with an amine of formula X ##STR19## wherein the symbols are as definedabove. The above-mentioned 3,3-bis-methyl-mercapto-2-cyano-acrylonitrileof formula IX is described under the name2,2-bis-methylmercapto-1-cyano-acrylonitrile' by R. Gompper and W.Topel, Chem. Ber. 95, 2861-2870, especially in the middle of page 2868,and can be prepared at a temperature of from 5 to 20° C. by the additionof malonic acid dinitrile having the formula CH₂ (CN)₂ to carbondisulfide in the presence of sodium methanolate in methanol, followed bymethylation of the resulting intermediate with dimethyl sulfate.

The starting material of formula III wherein X is the group (C[R₃]--R₄)_(q) wherein q is 0, and the remaining symbols are as definedabove, that is to say a compound of formula IIIa ##STR20## wherein R₂and n are as defined above, is obtained, for example, by reacting acompound of formula XI ##STR21## wherein R₂ and n are as defined above,with the tetracyano epoxide of formula XII ##STR22##

The compounds of formula XI are obtained, for example, from an aldehydeof formula XIII ##STR23## wherein R₂ and n are as defined above, whichis first converted using sulfur and morpholine into a compound offormula XIV ##STR24## wherein R₂ and n are as defined above, whichcompound is then converted using methyl iodide in acetone, followed byhydrogen sulfide in pyridine, into a compound of formula XI.

Process b):

The starting material of formula XVI can be used in the form of a salt,for example in the form of an acetate. The reaction is carried out informic acid at elevated temperature, preferably at 100-250° C., such asespecially at 200° C.

The starting material of formula XV is obtained from a compound offormula III by reaction with hydrazine in a suitable solvent, such as asuitable alkanol, such as preferably methanol, for example at the refluxtemperature.

Process c):

The reaction is carried out at elevated temperature, preferably at100-250° C., such as especially at 200° C., in the presence or, wherepossible, in the absence of a solvent, i.e. the formamide derivative offormula XVII can be used simultaneously as the solvent.

Process d):

The reaction is carried out at elevated temperature, preferably at50-180° C., such as especially at 120° C., in the presence or, wherepossible, in the absence of a solvent, i.e. the amine derivative offormula XVI can be used simultaneously as the solvent. When v is 0, theamine derivative of formula XVI is preferably used in the form of asalt, for example in the form of a hydrochloride. When v is 1, the aminederivative of formula XVI is preferably used in the form of the freeamine.

The starting material of formula XVIII is obtained from a compound offormula XV by reaction with a suitable dimethylformamide acetal, such asN,N-dimethylformamide diethyl acetal, in a suitable solvent, such as asuitable aromatic hydrocarbon, such as especially toluene, at elevatedtemperature, preferably at 50-180° C., such as especially under reflux.

Process e):

The Dimroth rearrangement is carried out at elevated temperature, forexample at 70-200° C., preferably at 80-150° C., for example underreflux, in a suitable water-containing solvent mixture, for example amixture of water and a suitable ether, such as a cyclic ether, such asdioxane, for example a dioxane/water mixture in a ratio by volume of1:1.

The imine of formula XIX is obtained, for example, from a compound offormula XV in two stages, as follows:

In the first stage, a compound of formula XV is reacted with orthoformicacid triethyl ester of formula HC(OC₂ H₅)₃ to form anethoxymethyleneamino compound of formula XX ##STR25## wherein thesymbols are as defined above.

The reaction is carried out at elevated temperature, preferably at50-180° C., such as especially at 120° C., the orthoformic acid triethylester serving simultaneously as the solvent. The ethanol formed by thereaction is continuously distilled off from the reaction mixture.

In the second stage, the resulting compound of formula XX is reactedwith an amine of formula XVI wherein v is 0 or 1 and the remainingsymbols are as defined above to form the desired imine of formula XIX.The reaction is carried out in a suitable solvent, such as a suitablealcohol, for example an alkanol, such as preferably ethanol, at elevatedtemperature, preferably at 50-180° C., such as especially at 70-120° C.,for example at the reflux temperature.

Alternatively, the imine of formula XIX is obtained directly from acompound of formula XVIII by reaction with an amine of formula XVI[similarly to process d)] in admixture with the end product of formulaI. That reaction is carried out in a suitable solvent, such as asuitable alcohol, for example an alkanol, such as preferably ethanol, atelevated temperature, preferably at 50-180° C., such as especially at70-120° C., for example at the reflux temperature.

Acid addition salts of compounds of formula I are obtained in a mannerknown per se, for example by treatment with an acid or a suitable anionexchange reagent.

Acid addition salts can be converted into the free compounds incustomary manner, for example by treatment with a suitable basic agent.

Mixtures of isomers can be separated into the individual isomers in amanner known per se, for example by fractional crystallization,chromatography, etc.

The invention relates especially to a process for the preparation of a4-phenylamino-1H-pyrazolo[3,4-d]pyrimidine derivative of formula Ia##STR26## wherein m and n are each independently of the other an integerfrom 0 up to and including 3,

R₁ is halogen, cyano, trifluoromethyl, hydroxy, lower alkoxy, loweralkanoyloxy, carboxy, lower alkoxycarbonyl, carbamoyl, N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by cyano, it being possible when several phenyl substituents R₁are present for those substituents to be identical or different from oneanother,

X is the group NH(CH₂)_(t) wherein t is an integer from 0 up to andincluding 3, or the group (C[R₃ ]--R₄)_(q) wherein q is an integer from0 up to and including 3, R₃ is hydrogen or lower alkyl and R₄ ishydrogen or lower alkyl, and

R₂ is halogen, nitro, cyano, trifluoromethyl, carboxy, loweralkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, amino, loweralkylamino, acylated amino having up to 10 carbon atoms, hydroxy, loweralkanoyloxy, oxa-lower alkoxy, lower alkoxy that is unsubstituted orsubstituted by carboxy, lower alkoxycarbonyl, carbamoyl or by N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by hydroxy, it being possible when several phenyl substituentsR₂ are present for those substituents to be identical or different fromone another,

or a salt thereof,

which process comprises treating a compound of formula IIa ##STR27##wherein R₅ is hydrogen or methyl, R₆ is alkoxy having from 1 to 3 carbonatoms or is nitro,

r is an integer from 0 to 2,

and the remaining substituents and symbols are as defined above, with asuitable Lewis acid and, if desired, converting an obtainable compoundof formula I into its salt, or converting an obtainable salt of acompound of formula I into the free compound.

The processes described above, including the processes for the removalof protecting groups and the additional process steps, are, unlessotherwise indicated, carried out in a manner known per se, for examplein the presence or absence of preferably inert solvents and diluents, ifnecessary in the presence of condensation agents or catalysts, atreduced or elevated temperature, for example in a temperature range offrom approximately -20° C. to approximately 200° C. (preferably to 150°C.), especially from approximately 0° C. to approximately 120° C.(preferably to +70° C. or to +80° C.), preferably from approximately+10° C. to approximately +50° C., above all at room temperature, in asuitable vessel and, if necessary, under an inert gas atmosphere, forexample a nitrogen atmosphere.

In those processes, taking into account all the substituents present inthe molecule, if necessary, for example when readily hydrolyzableradicals are present, especially mild reaction conditions should beused, such as short reaction times, the use of mild acidic or basicagents at low concentrations, stoichiometric quantity ratios and thechoice of suitable catalysts, solvents, temperature and/or pressureconditions.

The invention relates also to those forms of the process according towhich a compound obtainable as intermediate at any stage of the processis used as starting material and the remaining process steps are carriedout or the process is interrupted at any stage or a starting material isformed under the reaction conditions or is used in the form of areactive derivative or salt. There are preferably used those startingmaterials which in accordance with the process result in the compoundsdescribed above as being especially valuable.

The present invention relates also to novel starting materials and/orintermediates and to processes for the preparation thereof. The startingmaterials used and the reaction conditions chosen are preferably thosethat result in the compounds described in this Application as beingespecially preferred.

The intermediates of formulae V, VI, VII, XVIII, XIX and XX whereincorresponding substituents have the meanings mentioned in this text forthe compounds of the formula I are novel and the present inventionrelates also thereto. Preference is given to those intermediates thatresult in the preferred end products of formula I.

The invention relates to 5-amino-4-cyano-pyrazole derivatives of formulaV ##STR28## wherein n is an integer from 0 up to and including 3, r isan integer from 0 to 2,

X is the group NH(CH₂)_(t) wherein t is an integer from 0 up to andincluding 3, or the group (C[R₃ ]--R₄)_(q) wherein q is an integer from0 up to and including 3, R₃ is hydrogen or lower alkyl and R₄ ishydrogen or lower alkyl,

R₂ is halogen, nitro, cyano, tri-fluoromethyl, carboxy, loweralkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, amino, loweralkylamino, acylated amino having up to 10 carbon atoms, hydroxy, loweralkanoyloxy, oxa-lower alkoxy, lower alkoxy that is unsubstituted orsubstituted by carboxy, lower alkoxycarbonyl, carbamoyl or by N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by hydroxy, it being possible when several phenyl substituentsR₂ are present for those substituents to be identical or different fromone another,

R₅ is hydrogen or methyl, and

R₆ is alkoxy having from 1 to 3 carbon atoms or is nitro.

The invention relates also to compounds of formula XIX ##STR29## whereinm and n are each independently of the other an integer from 0 up to andincluding 3,

v is 0 or 1,

R is hydrogen or lower alkyl,

R₁ is halogen, cyano, trifluoromethyl, hydroxy, lower alkoxy, loweralkanoyloxy, carboxy, lower alkoxycarbonyl, carbamoyl, N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by cyano, it being possible when several phenyl substituents R₁are present for those substituents to be identical or different from oneanother,

X is the group NH(CH--R₇)_(t) wherein t is an integer from 0 up to andincluding 3 and R₇ is hydrogen or lower alkyl, or the group (C[R₃]--R₄)_(q) wherein q is an integer from 0 up to and including 3, R₃ ishydrogen or lower alkyl and R₄ is hydrogen or lower alkyl, and R₂ ishalogen, nitro, cyano, trifluoromethyl, carboxy, lower alkoxycarbonyl,carbamoyl, N-lower alkylcarbamoyl, N,N-di-lower alkylcarbamoyl, azido,amino, lower alkylamino, di-lower alkylamino, di-lower alkylamino-loweralkyleneamino, benzylamino; acylated or sulfonated amino each having upto 10 carbon atoms; hydroxy, lower alkanoyloxy, oxa-lower alkoxy, loweralkoxy that is unsubstituted or substituted by carboxy, loweralkoxycarbonyl, carbamoyl or by N-lower alkylcarbamoyl, or lower alkylthat is unsubstituted or substituted by amino, lower alkanoylamino,benzoylamino, lower alkoxycarbonylamino, sulfonated amino, cyano,hydroxy, lower alkanoyloxy, lower alkoxycarbonyloxy or by lower alkoxy,it being possible when several phenyl substituents R₂ are present forthose substituents to be identical or different from one another and fortwo vicinal radicals R₂ together also to form methylenedioxy,

and to salts of such compounds having salt-forming groups.

The compounds of the formula XIX can not only be used as intermediatesbut they exhibit also pharmacological activities similar to those of theend products of the formula I.

The invention relates also to 4-hydroxy-pyrazolo[3,4-d]pyrimidinederivatives of formula VI ##STR30## wherein n is an integer from 0 up toand including 3, r is an integer from 0 to 2,

X is the group NH(CH₂)_(t) wherein t is an integer from 0 up to andincluding 3, or the group (C[R₃ ]--R₄)_(q) wherein q is an integer from0 up to and including 3, R₃ is hydrogen or lower alkyl and R₄ ishydrogen or lower alkyl,

R₂ is halogen, nitro, cyano, trifluoromethyl, carboxy, loweralkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, amino, loweralkylamino, acylated amino having up to 10 carbon atoms, hydroxy, loweralkanoyloxy, oxa-lower alkoxy, lower alkoxy that is unsubstituted orsubstituted by carboxy, lower alkoxycarbonyl, carbamoyl or by N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by hydroxy, it being possible when several phenyl substituentsR₂ are present for those substituents to be identical or different fromone another,

R₅ is hydrogen or methyl, and

R₆ is alkoxy having from 1 to 3 carbon atoms or is nitro.

The invention relates also to 4-chloro-pyrazolo[3,4-d]pyrimidinederivatives of formula VII ##STR31## wherein n is an integer from 0 upto and including 3, r is an integer from 0 to 2,

X is the group NH(CH₂)_(t) wherein t is an integer from 0 up to andincluding 3, or the group (C[R₃ ]--R₄)_(q) wherein q is an integer from0 up to and including 3, R₃ is hydrogen or lower alkyl and R₄ ishydrogen or lower alkyl,

R₂ is halogen, nitro, cyano, trifluoromethyl, carboxy, loweralkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, amino, loweralkylamino, acylated amino having up to 10 carbon atoms, hydroxy, loweralkanoyloxy, oxa-lower alkoxy, lower alkoxy that is unsubstituted orsubstituted by carboxy, lower alkoxycarbonyl, carbamoyl or by N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by hydroxy, it being possible when several phenyl substituentsR₂ are present for those substituents to be identical or different fromone another,

R₅ is hydrogen or methyl, and

R₆ is alkoxy having from 1 to 3 carbon atoms or is nitro.

The invention relates also to compounds of formula XVIII ##STR32##wherein n is an integer from 0 up to and including 3, X is the groupNH(CH--R₇)_(t) wherein t is an integer from 0 up to and including 3 andR₇ is hydrogen or lower alkyl, or the group (C[R₃ ]--R₄)_(q) wherein qis an integer from 0 up to and including 3, R₃ is hydrogen or loweralkyl and R₄ is hydrogen or lower alkyl, and

R₂ is halogen, nitro, cyano, trifluoromethyl, carboxy, loweralkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, N,N-di-loweralkylcarbamoyl, azido, amino, lower alkylamino, di-lower alkylamino,di-lower alkylamino-lower alkyleneamino, benzylamino; acylated orsulfonated amino each having up to 10 carbon atoms; hydroxy, loweralkanoyloxy, oxa-lower alkoxy, lower alkoxy that is unsubstituted orsubstituted by carboxy, lower alkoxycarbonyl, carbamoyl or by N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino, lower alkanoylamino, benzoylamino, lower alkoxycarbonylamino,sulfonated amino, cyano, hydroxy, lower alkanoyloxy, loweralkoxycarbonyloxy or by lower alkoxy, it being possible when severalphenyl substituents R₂ are present for those substituents to beidentical or different from one another and for two vicinal radicals R₂together also to form methylenedioxy,

and to salts of such compounds having at least one salt-forming group.

The invention relates also to a method of treating warm-blooded animalssuffering from a tumor disease, which method comprises administering towarm-blooded animals requiring such treatment an effectivetumor-inhibiting amount of a compound of formula I or of apharmaceutically acceptable salt thereof. The invention relates also tothe use of a compound of formula I or of a pharmaceutically acceptablesalt thereof for inhibiting EGF-receptor-specific protein tyrosinekinase C in warm-blooded animals or in the preparation of pharmaceuticalcompositions for use in the therapeutic treatment of the human or animalbody. In such a treatment, a warm-blooded animal of approximately 70 kgbody weight will receive effective doses that depend on species, age,individual conditions, mode of administration and individual symptoms,for example daily doses of approximately from 5 to 5000 mg, especiallyfrom 200 to 2000 mg.

The invention relates also to pharmaceutical compositions that comprisean effective amount of active ingredient, especially an amount that iseffective in the prophylaxis or treatment of one of the above-mentioneddiseases, together with pharmaceutically acceptable carriers that aresuitable for topical, enteral, for example oral or rectal, or parenteraladministration and that may be inorganic or organic, solid or liquid.There are used for oral administration especially tablets or gelatincapsules that comprise the active ingredient together with diluents, forexample lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/orglycerol, and/or lubricants, for example siliceous earth, talc, stearicacid or salts thereof, such as magnesium or calcium stearate, and/orpolyethylene glycol. Tablets can also comprise binders, for examplemagnesium aluminum silicate, starches, such as corn, wheat or ricestarch, gelatin, methylcellulose, sodium carboxymethylcellulose and/orpolyvinylpyrrolidone, and, if desired, disintegrators, for examplestarches, agar, alginic acid or a salt thereof, for example sodiumalginate, and/or effervescent mixtures, or adsorbents, colorings,flavorings and sweeteners. The pharmacologically active compounds of thepresent invention can also be used in the form of parenterallyadministrable compositions or in the form of infusion solutions. Suchsolutions are preferably isotonic aqueous solutions or suspensionswhich, for example in the case of lyophilized compositions that comprisethe active ingredient on its own or together with a carrier, for examplemannitol, can be made up before use. The pharmaceutical compositions maybe sterilized and/or may comprise excipients, for example preservatives,stabilizers, wetting agents and/or emulsifiers, solubilizers, salts forregulating the osmotic pressure and/or buffers. The pharmaceuticalcompositions in question, which, if desired, may comprise furtherpharmacologically active substances, such as antibiotics, are preparedin a manner known per se, for example by means of conventional mixing,granulating, confectioning, dissolving or lyophilizing processes, andcomprise from approximately 0.1% to 100%, especially from approximately5% to approximately 90%, active ingredient or ingredients.

The Examples that follow illustrate the invention without limiting theinvention in any way. The ratio of solvents or eluants in the solvent oreluant mixtures used is given in parts by volume (v/v) and temperaturesare given in degrees Celsius.

Abbreviations:

abs.: absolute

APCI-MS: atmospheric pressure chemical ionisation mass spectrum

BOC: tert. butoxycarbonyl

DIPE: diisopropyl ether

DMEU: 1,3-dimethyl-2-imidazolidinone

DMF: dimethylformamide

EI-MS: electron impact ionization-mass spectroscopy

FAB-MS: fast atom bombardment-mass spectroscopy

HV: high vacuum

conc.: concentrated

min: minute(s)

RF: reflux

RT: room temperature

RV: rotary evaporator

brine: saturated sodium chloride solution

THF: tetrahydrofuran

TLC: thin layer chromatography

HPLC gradients:

    ______________________________________                                        Grad.sub.20-100                                                                              20% → 100% a) in b) for 20 min.                         Grad.sub.5-40   5% → 40% a) in b) for 20 min.                          ______________________________________                                    

Eluant a): acetonitrile +0.05% TFA; eluant b): water +0.05% TFA. Column(250×4.6 mm) filled with reversed-phase material C18-Nucleosil (5 maverage particle size, silica gel covalently derivatized withoctadecylsilanes, Macherey & Nagel, Duren, Germany). Detection by UVabsorption at 254 nm. The retention times (t_(Ret)) are given inminutes. Flow rate: 1 ml/min.

EXAMPLE 1

With the exclusion of air and moisture, there is added to a suspensionof 203 mg of AlCl₃ in 2.5 ml of absolute benzene a solution of 100 mg of1-benzyl-3,4-diphenylamino-pyrazolo[3,4-d]pyrimidine in 2.5 ml ofabsolute benzene. The reaction mixture is stirred for 1.5-2 hours at 50°C. until, according to thin-layer chromatography, no starting materialremains and is then stirred in approx. 30 ml of water. The precipitateis filtered off and dissolved in ethyl acetate. The ethyl acetate phaseis washed several times with 5% aqueous sodium hydrogen carbonatesolution and then with saturated sodium chloride solution, dried andconcentrated to dryness by evaporation. The residue is crystallized fromethyl acetate/hexane, yielding3,4-diphenylamino-1H-pyrazolo[3,4-d]pyrimidine in the form of colorlesscrystals, m.p. 263-264° C., FAB-MS: (M+H)⁺ =303 (C₁₇ H₁₄ N₆).

The starting material is obtained as follows:

Step 1.1:

With ice-cooling, 19.3 ml of a 5.4N sodium methanolate solution in 20 mlof methanol (purissimum) are added to 9.9 g of benzylhydrazinedihydrochloride in 20 ml of methanol (purissimum) and the reactionmixture is stirred for approx. 15 minutes at room temperature and thenintroduced into a solution of 4.65 g of2-cyano-3-methylmercapto-3-phenylamino-acrylonitrile in 150 ml ofabsolute ethanol. The mixture is heated under reflux for approx. 17hours and cooled to room temperature and insoluble material is filteredoff with suction. The filtrate is concentrated by evaporation using arotary evaporator and the brown oily residue is chromatographed on 190 gof silica gel, methylene chloride/ethyl acetate mixtures being used aseluant. 5-Amino-1-benzyl-4-cyano-3-phenylamino-pyrazole is obtained;m.p. 139-140° C., FAB-MS: (M+H)⁺ =290 (C₁₇ H₁₅ N₅).

Step 1.2:

1 g of 5-amino-1-benzyl-4-cyano-3-phenylamino-pyrazole and 6 ml of 85%aqueous formic acid are heated under reflux for 12 hours and then cooledto room temperature. The suspension is stirred with 20 ml of ethanol andthe crude product is filtered off with suction, made into a slurry withwater and again filtered off with suction. Recrystallized fromtetrahydrofuran/cyclohexane yields crystalline1-benzyl-4-hydroxy-3-phenylamino-pyrazolo[3,4-d]pyrimidine; m.p.246-247° C., FAB-MS: (M+H)⁺ =318 (C₁₈ H₁₅ N₅ O).

Step 1.3:

Under argon, 200 mg of1-benzyl-4-hydroxy-3-phenylamino-pyrazolo[3,4-d]-pyrimidine are heatedunder reflux for 5 hours with 2 ml of POCl₃, during which time thesuspension slowly becomes a solution. The light-brown solution is cooledto room temperature, concentrated to dryness by evaporation and stirredwith ice-water. The crude product is filtered off with suction andrecrystallized from ethanol/water, yielding fine needles of1-benzyl-4-chloro-3-phenylamino-pyrazolo[3,4-d]pyrimidine; m.p. 135° C.,FAB-MS: (M+H)⁺ =336 (C₁₈ H₁₄ ClN₅).

Step 1.4:

1 g of 1-benzyl-4-chloro-3-phenylamino-pyrazolo[3,4-d]pyrimidine issuspended in 8 ml of ethanol; 27 ml of aniline are added thereto and,under nitrogen, the reaction mixture is heated under reflux for 2.5hours until, according to thin-layer chromatography, all the startingmaterial has disappeared. The reaction mixture is concentrated todryness by evaporation, the residue is suspended in water and the pH isadjusted to pH 8.5-9 with 0.1N NaOH. Extraction is then carried out withethyl acetate and the ethyl acetate phase is then dried and concentratedby evaporation. The crude product is chromatographed on silica gel,mixtures of toluene/ethyl acetate being used as eluant.Product-containing column fractions are stirred with cyclohexane/hexane,yielding colorless crystals of1-benzyl-3,4-diphenylamino-pyrazolo[3,4-d]pyrimidine; FAB-MS: (M+H)⁺=393 (C₂₄ H₂₀ N₆).

EXAMPLE 2

In a manner analogous to that described in Example 1, there is obtainedfrom1-benzyl-3-(3-chloro-phenylamino)-4-phenylamino-pyrazolo[3,4-d]pyrimidine,by removal of the benzyl protecting group in AlCl₃ /benzene,3-(3-chloro-phenylamino)-4-phenylamino-1H-pyrazolo[3,4-d]pyrimidine;m.p. 230° C., FAB-MS: (M+H)⁺ =337 (C₁₇ H₁₃ ClN₆).

The starting material is obtained as follows:

Step 2.1:

Analogously to Step 1.1, there is obtained from3-(3-chloro-phenylamino)-2-cyano-3-methylmercapto-acrylonitrile andbenzylhydrazine dihydrochloride5-amino-1-benzyl-3-(3-chloro-phenylamino)-4-cyano-pyrazole; m.p.146-148° C.; FAB-MS: (M+H)⁺ =324 (C₁₇ H₁₄ ClN₅).

Step 2.2:

Analogously to Step 1.2, there is obtained from5-amino-1-benzyl-3-(3-chloro-phenylamino)-4-cyano-pyrazole, by boilingwith formic acid,1-benzyl-3-(3-chloro-phenylamino)-4-hydroxy-pyrazolo[3,4-d]pyrimidine;m.p. 234-236° C., FAB-MS: (M+H)⁺ =352 (C₁₈ H₁₄ ClN₅ O).

Step 2.3:

Analogously to Step 1.3, there is obtained from1-benzyl-3-(3-chloro-phenylamino)-4-hydroxy-pyrazolo[3,4-d]pyrimidine,by boiling for 7 hours in POCl₃ and crystallization from ethanol,1-benzyl-4-chloro-3-(3-chloro-phenylamino)-pyrazolo[3,4-d]pyrimidine;m.p. 148-150° C., FAB-MS: (M+H)⁺ =370 (C₁₈ H₁₃ Cl₂ N₅).

Step 2.4:

Analogously to Step 1.4, there is obtained from1-benzyl-4-chloro-3-(3-chloro-phenylamino)-pyrazolo[3,4-d]pyrimidine andaniline, by boiling in ethanol,1-benzyl-3-(3-chloro-phenylamino)-4-phenylamino-pyrazolo[3,4-d]pyrimidine;m.p. 64-65° C.; FAB-MS: (M+H)⁺ =427 (C₂₄ H₁₉ ClN₆).

EXAMPLE 3

Analogously to Example 1, there is obtained from1-benzyl-3,4-di(3-chloro-phenylamino)pyrazolo[3,4-d]pyrimidine, byremoval of the benzyl protecting group in AlCl₃ /benzene,3,4-di(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine; m.p. 1 75°C., FAB-MS: (M+H)⁺ =371 (C₁₇ H₁₂ Cl₂ N₆).

The starting material is obtained as follows:

Step 3.1:

Analogously to Step 1.4, there is obtained from1-benzyl-4-chloro-3-(3-chloro-phenylamino)-pyrazolo[3,4-d]pyrimidine(see Step 2.3) and 3-chloro-aniline, by boiling in ethanol,1-benzyl-3,4-di(3-chloro-phenylamino)pyrazolo[3,4-d]pyrimidine; m.p.131-133° C., FAB-MS: (M+H)⁺ =461 (C₂₄ H₁₈ Cl₂ N₆).

EXAMPLE 4

Analogously to Example 1, there is obtained from1-benzyl-4-(3-bromo-phenylamino)-3-(3-chloro-phenylamino)-pyrazolo[3,4-d]pyrimidine,by removal of the benzyl protecting group in AlCl₃ benzene,4-(3-bromo-phenylamino)-3-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;m.p. 179-181° C., FAB-MS: (M+H)⁺ =415 (C₁₇ H₁₂ BrClN₆).

The starting material is obtained as follows:

Step 4.1:

Analogously to Step 1.4, there is obtained from1-benzyl-4-chloro-3-(3-chloro-phenylamino)-pyrazolo[3,4-d]pyrimidine(see Step 2.3) and 3-bromo-aniline, by boiling in ethanol,1-benzyl-4-(3-bromo-phenylamino)-3-(3-chloro-phenylamino)-pyrazolo[3,4-d]-pyrimidine;FAB-MS: (M+H)⁺ =506 (C₂₄ H₁₈ ClBrN₆).

EXAMPLE 5

Analogously to Example 1, there is obtained from1-benzyl-3-(3-chloro-phenylamino)-4-(3-methyl-phenylamino)-pyrazolo[3,4-d]pyrimidine,by removal of the benzyl protecting group in AlCl₃ /benzene,3-(3-chloro-phenylamino)-4-(3-methyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;m.p. 194-195° C., FAB-MS: (M+H)⁺ =351 (C₁₈ H₁₅ ClN₆).

The starting material is obtained as follows:

Step 5.1:

Analogously to Step 1.4, there is obtained from1-benzyl-4-chloro-3-(3-chloro-phenylamino)-pyrazolo[3,4-d]pyrimidine(see Step 2.3) and 3-methyl-aniline, by boiling in ethanol,1-benzyl-3-(3-chloro-phenylamino)-4-(3-methyl-phenylamino)-pyrazolo[3,4-d]-pyrimidine;FAB-MS: (M+H)⁺ =441 (C₂₅ H₂₁ ClN₆).

EXAMPLE 6

Analogously to Example 1, there is obtained from1-benzyl-4-(3-[2-cyano-ethyl]-phenylamino)-3-phenylamino-pyrazolo[3,4-d]pyrimidine,by removal of the benzyl protecting group in AlCl₃ /benzene,4-(3-[2-cyano-ethyl]-phenylamino)-3-phenylamino-1H-pyrazolo-[3,4-d]pyrimidine;m.p. 202° C., FAB-MS: (M+H)⁺ =356 (C₂₀ H₁₇ N₇).

The starting material is obtained as follows:

Step 6.1:

Analogously to Step 1.4, there is obtained from1-benzyl-4-chloro-3-phenylamino-pyrazolo[3,4-d]pyrimidine (see Step 1.3)and 3-[2-cyano-ethyl]-aniline, by boiling in ethanol,1-benzyl-4-(3-[2-cyano-ethyl]-phenylamino)-3-phenylamino-pyrazolo[3,4-d]pyrimidine;FAB-MS: (M+H)⁺ =446 (C₂₇ H₂₃ N₇).

EXAMPLE 7

Analogously to Example 1, there is obtained from1-benzyl-4-(4-[2-cyano-ethyl]-phenylamino)-3-phenylamino-pyrazolo[3,4-d]pyrimidine,by removal of the benzyl protecting group in AlCl₃ /benzene,4-(4-[2-cyano-ethyl]-phenylamino)-3-phenylamino-1H-pyrazolo-[3,4-d]pyrimidine;m.p. 268° C.; FAB-MS: (M+H)⁺ =356 (C₂₀ H₁₇ N₇).

The starting material is obtained as follows:

Step 7.1:

Analogously to Step 1.4, there is obtained from1-benzyl-4-chloro-3-phenylamino-pyrazolo[3,4-d]pyrimidine (see Step 1.3)and 4-[2-cyano-ethyl]-aniline, by boiling in ethanol,1-benzyl-4-(4-[2-cyano-ethyl]-phenylamino)-3-phenylamino-pyrazolo[3,4-d]pyrimidine;FAB-MS: (M+H)⁺ =446 (C₂₇ H₂₃ N₇).

EXAMPLE 8

Analogously to Example 1, there is obtained from1-benzyl-4-(3-cyanomethyl-phenylamino)-3-phenylamino-pyrazolo[3,4-d]pyrimidine,by removal of the benzyl protecting group in AlCl₃ /benzene,4-(3-cyanomethyl-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]-pyrimidine.

The starting material is obtained as follows:

Step 8.1:

Analogously to Step 1.4, there is obtained from1-benzyl-4-chloro-3-phenylamino-pyrazolo[3,4-d]pyrimidine (see Step 1.3)and 3-cyanomethyl-aniline, by boiling in ethanol,1-benzyl-4-(3-cyanomethyl-phenylamino)-3-phenylamino-pyrazolo[3,4-d]pyrimidine;m.p. 80° C., FAB-MS: (M+H)⁺ =432 (C₂₆ H₂₁ N₇).

EXAMPLE 9

1.3 g of3-(3-chloro-phenylamino)-4-(3-methyl-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine(see Example 5) are dissolved in 70 ml of ethanol and at roomtemperature 2 ml of a 4N solution of hydrogen chloride in ethanol areadded thereto. After approximately 10 minutes' stirring, the HCl saltbegins to crystallize out. The solution is cooled to 0° C. and the saltis crystallized out completely by the addition of ethyl ether, yieldingcolorless crystals of3-(3-chloro-phenylamino)-4-(3-methyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidinehydrochloride; m.p. 260-263° C.

EXAMPLE 10

100 mg of4-(3-[2-cyano-ethyl]-phenylamino)-3-phenylamino-1H-pyrazolo-[3,4-d]pyrimidine(see Example 6) are dissolved in 2.5 ml of absolute tetrahydrofuran andadded dropwise in the course of 10 minutes to a suspension of 135 mg ofaluminum trichloride in 2.5 ml of tetrahydrofuran and 46 mg of lithiumaluminum hydride. The reaction is slightly exothermic. The reactionmixture is heated for 2.5 hours under reflux until, according tothin-layer chromatography, no starting material remains. The solution iscooled to 0° C., 5 ml of water are added thereto and stirring is carriedout for 2 hours at room temperature. The pH is then adjusted to pH 9with 1N sodium hydroxide solution, insoluble material is filtered offand the filtrate is concentrated by evaporation. The residue is digestedin tetrahydrofuran. Again, insoluble material is filtered off. Thetetrahydrofuran filtrate is concentrated to approximately 3 ml andapprox. 15 ml of methylene chloride are added thereto. 5 ml of hexaneare added and at 0° C. crystals of4-(3-[3-amino-propyl]-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidineprecipitate.

EXAMPLE 11

There is obtained from4-(4-[2-cyano-ethyl]-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidine(see Example 7), by reduction with Raney nickel in a solution of ammoniain methanol and subsequent treatment with hydrogen chloride in ethanol,4-(4-[3-amino-propyl]-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidinehydrochloride.

EXAMPLE 12

There is obtained from4-(3-cyanomethyl-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidine(see Example 8), by reduction with Raney nickel in a solution of ammoniain methanol,4-(3-[2-amino-ethyl]-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidine.

EXAMPLE 13

The following compounds of formula I are obtained analogously to themethods described in this text, for example analogously to Examples 1-3:

a)3-(4-chloro-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,

b)4-(3-chloro-phenylamino)-3-(3-fluoro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,

c)4-(3-chloro-phenylamino)-3-(4-fluoro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,

d)4-(3-chloro-phenylamino)-3-(4-methoxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,

e)4-(3-chloro-phenylamino)-3-(4-hydroxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidineand

f)4-(3-chloro-phenylamino)-3-(4-iodo-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,for example starting from the following compounds of formula V:

aa) Analogously to Step 1.1, there is obtained from3-(4-chloro-phenylamino)-2-cyano-3-methylmercapto-acrylonitrile andbenzylhydrazine dihydrochloride5-amino-1-benzyl-3-(4-chloro-phenylamino)-4-cyano-pyrazole; m.p.163-164° C., FAB-MS (M+H)⁺ : 324 (C₁₇ H₁₄ ClN₅).

ba) Analogously to Step 1.1, there is obtained from2-cyano-3-(3-fluoro-phenylamino)-3-methylmercapto acrylonitrile andbenzylhydrazine dihydrochoride5-amino-1-benzyl-4-cyano-3-(3-fluoro-phenylamino)-pyrazole; m.p.151-152° C., FAB-MS (M+H)⁺ : 308 (C₁₇ H₁₄ FN₅).

ca) Analogously to Step 1.1, there is obtained from2-cyano-3-(4-fluoro-phenylamino)-3-methylmercapto-acrylonitrile andbenzylhydrazine dihydrochloride5-amino-1-benzyl-4-cyano-3-(4-fluoro-phenylamino)-pyrazole; m.p.167-168° C., FAB-MS (M+H)⁺ : 308 (C₁₇ H₁₄ FN₅).

da) Analogously to Step 1.1, there is obtained from2-cyano-3-(4-methoxy-phenylamino)-3-methylmercapto-acrylonitrile andbenzylhydrazine dihydrochloride5-amino-1-benzyl-4-cyano-3-(4-methoxy-phenylamino)-pyrazole; yellowresin, TLC: R_(f) =0.30 (acetic acid ethylate/hexane 1:1).

ea) Analogously to Step 1.1, there is obtained from2-cyano-3-(4-iodo-phenylamino)-3-methylmercapto-acrylonitrile andbenzylhydrazine dihydrochloride5-amino-1-benzyl-4-cyano-3-(4-iodo-phenylamino)-pyrazole; FAB-MS (M+H)⁺: 416 (C₁₇ H₁₄ IN₅).

EXAMPLE 14

With the exclusion of moisture, 79.2 g (295 mmol) ofN'-(3-benzylamino-4-cyano-1H-pyrazol-5-yl)-N,N-dimethylformamidine aresuspended in 700 ml of methanol; 60.6 g (369 mmol) of 3-chloro-anilinehydrochloride are added and the reaction mixture is boiled under refluxfor 22 hours. The resulting yellow reaction solution is cooled to ≈50°C. and poured onto 2 liters of ice-water, 200 ml of saturated NaHCO₃solution and 1 liter of ethyl acetate. The aqueous phase is separatedoff and extracted twice with ethyl acetate. The organic phases arewashed twice with water, saturated NaHCO₃ solution, water and brine,dried (Na₂ SO₄) and concentrated by evaporation to a residual volume of≈1.5 liters. Inoculation and dilution with 300 ml of diethyl ether yieldcrystalline3-benzylamino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;m.p. 214-217° C.; TLC: R_(f) =0.29 (ethyl acetate:hexane=1:1).

The starting material is prepared as follows:

Step 14.1:

43.6 ml (400 mmol) of benzylamine are added to a suspension of 68.4 g(400 mmol) of 3,3-bis-methylmercapto-2-cyano-acrylonitrile[3,3-bis(methylsulfanyl)-2-cyano-acrylonitrile] (Maybridge) in 400 ml ofethyl acetate. The clear solution is heated slowly to 70° C. (→evolution of MeSH!), stirred at that temperature for 1.5 hours, cooledto RT and concentrated by evaporation, yielding crystalline3-benzylamino-3-methylmercapto-2-cyano-acrylonitrile; ¹ H-NMR: (CD₃ OD)7.36 (m, 5H), 4.77 (s, 2H), 2.59 (s, 3H).

Step 14.2:

24 ml (0.48 mol) of hydrazine hydrate are added dropwise to a solutionof 92 g (0.4 mol) of3-benzylamino-3-methylmercapto-2-cyano-acrylonitrile in 400 ml ofmethanol, during which the temperature rises to 40° C. The reactionmixture is heated slowly to boiling (→ evolution of MeSH!), boiled for 2hours, cooled to RT and concentrated by evaporation to a residual volumeof ≈200 ml. Dilution with diethyl ether, filtering and washing withdiethyl ether yield 5-amino-3-benzylamino-1H-pyrazole-4-carbonitrile[Spectrochimica Acta, 47A, 1635 (1991)]; m.p. 150-152° C.; TLC: R_(f)=0.41 (ethyl acetate).

Step 14.3:

Under a N₂ atmosphere, a suspension of 74.3 g (348 mmol) of5-amino-3-benzylamino-1H-pyrazole-4-carbonitrile in 1.0 liter of tolueneis boiled under reflux for 2 hours with 70.1 ml (95%; 409 mmol) ofN,N-dimethylformamide diethyl acetal. Cooling to RT, filtering withsuction and washing with diethyl ether yieldN'-(3-benzylamino-4-cyano-1H-pyrazol-5-yl)-N,N-dimethylformamidine; m.p.197-200° C.; TLC: R_(f) =0.50 (ethyl acetate).

Step 14.4:

60 g (0.47 mol) of 3-chloro-aniline are dissolved in 255 ml (0.56 mol)of a 2.2N solution of HCl in methanol. Concentration and stirring of theresidue in diethyl ether, followed by filtering and drying, yield3-chloro-aniline hydrochloride.

EXAMPLE 15

788 mg (4.8 mmol) of 4-formyl-benzoic acid methyl ester and 300 mg of 5%Pt/C are added to a solution of 1.04 g (4.0 mmol) of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 40 mlof DMEU and 961 mg (16 mmol) of acetic acid and hydrogenation is carriedout immediately (instead of hydrogenation, reduction with NaCNBH₃ isalso possible). After two days and after four days, a further 788 mg of4-formylbenzoic acid methyl ester is added. After 7 days the catalyst isseparated off from the reaction mixture by filtration through Celite,the filtrate is decolorized by treatment with activated carbon and thenconcentrated by evaporation under a high vacuum at 70° C. to give aresidue of ≈4 g. Crystallization with diisopropyl ether/toluene yieldsproduct of ≈80% purity. Heating in a mixture of 30 ml of ethanol and ≈5ml of acetone, filtering while hot, concentration by evaporation to halfthe volume and cooling yield4-(3-chloro-phenylamino)-3-(4-methoxy-carbonylbenzylamino)-1H-pyrazolo[3,4-d]pyrimidine;m.p. 228° C.; FAB-MS: (M+H)⁺ =409.

The starting material is obtained as follows:

Step 15.1:

In order to remove residual water, some solvent is distilled off from asuspension of 75.8 g (216 mmol) of3-benzylamino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine in1.5 liters of benzene. Then, with exclusion of moisture, the suspensionis introduced into 84 g of aluminum chloride (Fluka, Buchs/Switzerland)in 500 ml of benzene. The reaction mixture is heated at 80° C. for 2.5hours and then cooled to RT. The supernatant benzene phase is pouredinto 2 kg of ice-water (leaving behind a green, oily residue) and thesolid that precipitates is filtered with suction and rinsed thoroughlywith water (→K₁). Using a rotary evaporator, the benzene is evaporatedoff from the filtrate, and the aqueous phase that remains is added,together with 1 kg of ice, to the green, oily residue and hydrolysis iscarried out for 2 hours at 40° C. The crystalline product is filteredwith suction and rinsed with water (→K₂). K₁ and K₂ are taken up in 1liter of methanol, acidified with 4N aqueous HCl and partiallyconcentrated by evaporation. Water is added and the methanol isevaporated off completely. The crystals are filtered off and rinsed withwater. The same purification procedure is repeated with semi-saturatedNa₂ CO₃ solution/methanol and water/methanol. Stirring at 50° C. inmethanol, precipitation with diethyl ether, filtering and drying yield3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine; m.p.232-234° C.; TLC: R_(f) =0.50 (ethyl acetate).

EXAMPLE 16

16.5 mg (0.39 mmol) of LiOH×H₂ O are added to a mixture of 98 mg (0.24mmol) of3-(4-methoxycarbonylbenzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidinein 8 ml of methanol and 4 ml of water and the reaction mixture isstirred for 3 days at 45° C. The reaction mixture is concentrated byevaporation and the residue is taken up in ethanol. After the additionof activated carbon, the reaction mixture is filtered until clear andthen concentrated by evaporation again. Precipitation from a solution inDMF with diethyl ether yields the lithium salt of3-(4-carboxy-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=9.7; FAB-MS: (M+H, acid)⁺ =395, (M+Li)⁺ =401.

EXAMPLE 17

A solution of 261 mg (1.0 mmol) of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine (see Step15.1) and 245 mg (1.5 mmol) of 3-(methylamino-carbonyl)-benzaldehyde in26 ml of methanol, 26 ml of DMEU and 180 mg (3.0 mmol) of acetic acid isstirred for 1 hour at RT. Then 440 mg (7.0 mmol) of NaCNBH₃ are addedand the reaction mixture is stirred for 9 days at RT. The methanol isevaporated off from the reaction solution using a rotary evaporator, theresidue is poured into 0.6 liter of water and stirring is carried outovernight. The product precipitates. Filtering with suction, washingwith water, stirring twice in 10 ml of boiling ethanol, cooling andfiltering yield4-(3-chloro-phenylamino)-3-[3-(methylaminocarbonyl)-benzylamino]-1H-pyrazolo[3,4-d]pyrimidine;m.p. 252-254° C.; HPLC: T_(Ret) (Grad₅₋₄₀)=16.9; FAB-MS: (M+H)⁺ =408.

The starting material is prepared as follows:

Step 17.1:

A solution of 3 g of 3-formyl-benzoic acid methyl ester and 20 ml ofmethylamine (8.03M in ethanol) is stirred for 3 days at RT.Concentration by evaporation and crystallization from DIPE yieldN-methyl-3-methyliminomethyl-benzamide; m.p. 87° C.

Step 17.2:

A two-phase mixture of 2.91 g of N-methyl-3-methyliminomethyl-benzamide,50 ml of methylene chloride and 30 ml of 1N HCl is stirred for 1.5 hoursat RT. Separating off the organic phase, washing with 1N HCl and brine,drying with Na₂ SO₄, concentrating by evaporation and stirring withDIPE/hexane yield 3-(methylaminocarbonyl)-benzaldehyde; m.p. 101-102° C.

EXAMPLE 18

Analogously to Example 17, there is prepared from 261 mg (1.0 mmol) of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine (see Step15.1) and 245 mg (1.5 mmol) of 4-(methylaminocarbonyl)-benzaldehyde,4-(3-chloro-phenylamino)-3-[4-(methylaminocarbonyl)-benzylamino]-1H-pyrazolo[3,4-d]pyrimidine;HPLC: T_(Ret) (Grad₅₋₄₀)=16.3; ¹ H-NMR: (DMSO-d₆) 12.38 (s, HN), 8.93(s, HN), 8.39 (m, 1H), 8.27 (s, 1H), 7.94 (t, J=2, 1H), 7.81 (d, J=8,2H), 7.68 (db, J=8, 1H), 7.52 (d, J=8, 2H), 7.41 (t, J=8, 1H), 7.17 (db,J=8, 1H), 6.99 (tb, J=5, HN), 4.57 (d, J=5, 2H), 2.79 (d, J=4, 3H);FAB-MS: (M+H)⁺ =408.

The starting material is prepared analogously to Steps 17.1 and 17.2:

Step 18.1:

There is obtained from 3 g of 4-formyl-benzoic acid methyl ester and 20ml of methylamine (8.03M in ethanol)N-methyl-4-methyliminomethyl-benzamide; m.p. 140-141° C.

Step 18.2:

705 mg of N-methyl-4-methyliminomethyl-benzamide are hydrolyzed to form3-(methylaminocarbonyl)-benzaldehyde; ¹ H-NMR: (CDCl₃) 10.06 (s, 1H),7.93 (s, 4H), 6.4 (sb, HN), 3.04 (d, J=5, 3H).

EXAMPLE 19

99.7 mg (0.60 mmol) of 3,5-dimethoxy-benzaldehyde are added to asolution of 130 mg (0.50 mmol) of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol and 120 mg (2.0 mmol) of acetic acid. As the reactionmixture is stirred, a solid precipitates which can be dissolved again bythe addition of 52 ml of DMEU. 220 mg (3.5 mmol) of NaCNBH₃ are addedthereto and stirring is then continued for 5 hours at RT. Since not allof the 3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine hasbeen reacted (HPLC), 220 mg of NaCNBH₃ are added twice more and stirringis continued for 5 hours each time. The reaction solution is then pouredinto 1 liter of water, stirred vigorously for 1 hour and filtered.Crystallization of the filtration residue from acetone yields4-(3-chloro-phenylamino)-3-(3,5-dimethoxybenzylamino)-1H-pyrazolo[3,4-d]pyrimidine;m.p. 208-209° C.; HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=11.7; FAB-MS: (M+H)⁺ =411.

EXAMPLE 20

228 mg (1.50 mmol) of 4-hydroxy-3-methoxy-benzaldehyde are added (N₂atmosphere) to a solution of 261 mg (1.00 mmol) of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 26 ml of DMEU and 120 mg (2.0 mmol) of acetic acid. After 1hour, 440 mg (7.0 mmol) of NaCNBH₃ are added to the clear solution andstirring is then carried out at RT for 7 days. The reaction solution ispoured into 0.8 liter of water and stirred overnight to complete thereaction, during which time the product precipitates. Filtering withsuction, washing with water, stirring in hot ethyl acetate, cooling andfiltering yield4-(3-chloro-phenylamino)-3-[(3-methoxy-4-hydroxy-benzyl)-amino]-1H-pyrazolo[3,4-d]pyrimidine;m.p. 223-225° C.; HPLC: T_(Ret) (Grad₅₋₄₀)=19.8; FAB-MS: (M+H)⁺ =397.

EXAMPLE 21

225 mg (1.50 mmol) of 3-formyl-benzoic acid are added to a solution of261 mg (1.00 mmol) of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 120 mg (2.0 mmol) of acetic acid under aN₂ atmosphere. The reaction mixture is stirred for 1 hour, during whichtime a solid precipitates, and then 440 mg (7.0 mmol) of NaCNBH₃ areadded. Stirring is carried out for 5 days, the suspension changing to aclear solution. The methanol is evaporated off using a rotaryevaporator. The residue is poured into 0.6 liter of water and stirredfor 3 hours to complete the reaction. Filtering with suction, washingwith water and diethyl ether, stirring in hot ethanol, cooling andfiltering yield3-(3-carboxy-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine(3-{[4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidin-3-yl-amino]methyl}-benzoicacid); m.p. 294-296° C.; HPLC: T_(Ret) (Grad₅₋₄₀)=20.1; FAB-MS: (M+H)⁺=395.

EXAMPLE 22

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 3-formyl-benzoic acid methyl ester and then reduced with 7.00 mmolof NaCNBH₃ (5-7 days).4-(3-Chloro-phenylamino)-3-(3-methoxycarbonyl-benzylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained.

Preferably, the same compound is obtained as follows: To 70 mg (0.177mmol) of3-(3-carboxy-benzyl-amino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine(cf. Example 21) and 2 ml of methanol are added 0.1 ml ofthionylchloride. The mixture is stirred in a sealed vessel for 3 h at70° C., and then cooled to room temperature. Filtration and washing withmethanol affords4-(3-chloro-phenylamino)-3-(3-methoxycarbonyl-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine-hydrochloride;(C₂₀ H₁₇ N₆ ClO₂ ×HCl×0.28H₂ O), calc. C 53.34, H 4.15, N 18.66, Cl15.74, H₂ O 1.12, found C 53.35, H 4.13, N 18.82, Cl 15.87, H₂ O 1.12.

EXAMPLE 23

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 3,4,5-trimethoxy-benzaldehyde and then reduced with 7.00 mmol ofNaCNBH₃ (5-7 days).4-(3-Chloro-phenylamino)-3-(3,4,5-trimethoxybenzylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained; m.p. 240-245° C. HPLC: T_(Ret) (Grad₅₋₄₀)=22.

EXAMPLE 24

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 3,4-dimethoxy-benzaldehyde and then reduced with 7.00 mmol ofNaCNBH₃ (5-7 days).4-(3-Chloro-phenylamino)-3-(3,4-dimethoxybenzylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained; m.p. 228-232° C.; T_(Ret) (Grad₅₋₄₀)=19.0.

EXAMPLE 25

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 2,3,4-trimethoxy-benzaldehyde and then reduced with 7.00 mmol ofNaCNBH₃ (5-7 days).4-(3-Chloro-phenylamino)-3-(2,3,4-trimethoxybenzylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained; m.p. 168-169° C.; HPLC: T_(Ret) (Grad₅₋₄₀)=20.2.

EXAMPLE 26

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 3-hydroxy-4-methoxy-benzaldehyde and then reduced with 7.00 mmol ofNaCNBH₃ (5-7 days).4-(3-Chloro-phenylamino)-3-(3-hydroxy-4-methoxy-benzylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained; m.p. 225-227° C.; HPLC: T_(Ret) (Grad₅₋₄₀)=17.6.

EXAMPLE 27

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 4-hydroxy-3,5-dimethoxy-benzaldehyde and then reduced with 7.00mmol of NaCNBH₃ (5-7 days).4-(3-Chloro-phenylamino)-3-(4-hydroxy-3,5-dimethoxy-benzylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained; HPLC: T_(Ret) (Grad₅₋₄₀)=17.0.

EXAMPLE 28

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 3,4-methylenedioxy-benzaldehyde and then reduced with 7.00 mmol ofNaCNBH₃ (5-7 days).4-(3-Chloro-phenylamino)-3-(3,4-methylenedioxybenzylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained; m.p. 220-222° C.; HPLC: T_(Ret) (Grad₅₋₄₀)=22.5° C.

EXAMPLE 29

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 2,3-methylenedioxy-benzaldehyde and then reduced with 7.00 mmol ofNaCNBH₃ (5-7 days).4-(3-Chloro-phenylamino)-3-(2,3-methylenedioxybenzylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained; m.p. 214-215° C.; HPLC: T_(Ret) (Grad₅₋₄₀)=24.1.

EXAMPLE 30

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 3-chloro-benzaldehyde and then reduced with 7.00 mmol of NaCNBH₃(5-7 days).3-(3-Chloro-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo-[3,4-d]pyrimidineis obtained; m.p. 158-163° C.; HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=12.4.

EXAMPLE 31

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 3-chloro-4-hydroxy-benzaldehyde and then reduced with 7.00 mmol ofNaCNBH₃ (5-7 days).3-(3-Chloro-4-hydroxy-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained; m.p. 220° C.; HPLC: T_(Ret) (Grad₅₋₄₀)=18.7.

EXAMPLE 32

Analogously to Example 21, 1.00 mmol of3-amino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine in 26 mlof methanol, 13 ml of DMEU and 3.0 mmol of acetic acid are first reactedwith 3-chloro-4-methoxy-benzaldehyde and then reduced with 7.00 mmol ofNaCNBH₃ (5-7 days).3-(3-Chloro-4-methoxy-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidineis obtained; m.p. 205-208° C.; HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=12.2.

EXAMPLE 33

There are obtained analogously to the processes described in this text:

a)4-(3-chloro-phenylamino)-3-{[1-(3-chloro-phenyl)-ethyl]-amino}-1H-pyrazolo[3,4-d]-pyrimidine,

b)4-(3-chloro-phenylamino)-3-[(1-phenyl-ethyl)-amino]-1H-pyrazolo[3,4-d]pyrimidine[prepared as described in example 14, starting from(±)-1-phenyl-ethylamine and3,3-bis-(methylmercapto)-2-cyano-acrylonitril]; T_(Ret)(Grad₂₀₋₁₀₀)=11.8; FAB-MS: (M+H)⁺ =365, and

c)4-(3-chloro-phenylamino)-3-[3-(dimethylaminocarbonyl)-benzylamino]-1H-pyrazolo[3,4-d]-pyrimidine.

EXAMPLE 34

1.31 g (7.98 mmol) of 3-chloro-aniline hydrochloride (see Step 14.4) areadded to a suspension of 2.05 g (7.6 mmol) ofN'-[3-(4-methoxy-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethyl-formamidinein 24 ml of methanol and the reaction mixture is boiled under reflux.After 13 hours a further 561 mg (3.42 mmol) of 3-chloro-anilinehydrochloride is added. After a total boiling time of 18 hours, thesuspension is cooled.4-(3-Chloro-phenylamino)-3-(4-methoxy-phenyl)-1H-pyrazolo[3,4-d]pyrimidineis filtered off, washed with hexane and dried: m.p. 268-269° C.; HPLC:T_(Ret) (Grad₂₀₋₁₀₀)=13.8; FAB-MS: (M+H)⁺ =352.

The starting material is prepared as follows:

Step 34.1:

Under a N₂ atmosphere, a suspension of 1.87 g (8.16 mmol) of5-amino-3-(4-methoxy-phenyl)-1H-pyrazole-4-carbonitrile (for preparationsee: J. Heterocyclic Chem. 27, 647 (1990)) in 33 ml of toluene is boiledunder reflux for 3.5 hours with 1.64 ml of N,N-dimethylformamide diethylacetal (95%; 9.1 mmol). Crystallization by cooling to 0° C., filteringwith suction and washing with hexane yieldN'-[3-(4-methoxy-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidine;m.p. 169-171° C.; TLC: R_(f) =0.18 (ethyl acetate:hexane=1:1); MS: (M)⁺=269.

EXAMPLE 35

With the exclusion of moisture, 62 mg (0.38 mmol) of 3-chloro-anilinehydrochloride (see Step 14.4) are added to 92 mg (0.36 mmol) ofN'-[3-(4-amino-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidinein 1 ml of methanol and the reaction mixture is boiled under reflux for28 hours. The light-yellow reaction solution is cooled, concentrated byevaporation and chromatographed (SiO₂, methylene chloride/ethanol[10:1]). Crystallization from isopropanol yields3-(4-amino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;TLC: R_(f) =0.33 (methylene chloride:methanol=10:1); FAB-MS: (M)⁺ =336;HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=9.2, T_(Ret) (Grad₅₋₄₀)=18.8.

As an alternative to the above method,3-(4-amino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidinecan also be prepared from3-(4-tert-butoxycarbonylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine(Example 36) by stirring for 3 hours in 4N HCl solution in dioxane andfiltering off the product in the form of the hydrochloride.

The starting material is prepared as follows:

Step 35.1:

Under a N₂ atmosphere, 0.92 g (6.38 mmol) of tetracyanoethylene oxide(Fluka, Buchs/Switzerland) is added to 618.5 mg (2.9 mmol) of4-nitro-dithiobenzoic acid methyl ester (for preparation see J. prakt.Chem. 331, 243 (1989)) in 5 ml of toluene and the reaction mixture isthen boiled for 4 hours. The reaction mixture is cooled, 10 g of silicagel are added thereto and the reaction mixture is concentrated byevaporation using a rotary evaporator. Application of the residue to asilica gel column and elution with hexane/ethyl acetate (2:1) yield3-(4-nitrophenyl)-3-methylmercapto-2-cyano-acrylonitrile; TLC: R_(f)=0.46 (ethyl acetate:hexane=1:2); MS: (M)⁺ =245.

Step 35.2:

0.05 ml (1.00 mmol) of hydrazine hydrate is added dropwise to 245.3 mg(1.00 mmol) of 3-(4-nitrophenyl)-3-methylmercapto-2-cyano-acrylonitrilein 1.3 ml of methanol and the reaction mixture is then boiled for 8hours and then concentrated by evaporation. The residue is stirred withethyl acetate and filtered, yielding5-amino-3-(4-nitrophenyl)-1H-pyrazole-4-carbonitrile; TLC: R_(f) =0.14(ethyl acetate:hexane=1:1).

Step 35.3:

In 1 ml of toluene, 57.3 mg (0.25 mmol) of5-amino-3-(4-nitro-phenyl)-1H-pyrazole-4-carbonitrile and 79.3 μl ofN,N-dimethylformamide dibenzylacetal are boiled overnight. Filtering ofthe suspension and washing with hexane yieldN'-[3-(4-nitro-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidine;TLC: R_(f) =0.15 (ethyl acetate:hexane=2:1); T_(Ret) (Grad₂₀₋₁₀₀)=8.8.

Step 35.4:

In the presence of 30 mg of Pd/C (5%), 142 mg (0.50 mmol) ofN'-[3-(4-nitro-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidineare hydrogenated in 20 ml of THF. Filtering, concentration byevaporation and crystallization from ethyl acetate/diethyl ether/hexaneyieldN'-[3-(4-amino-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidine;TLC: R_(f) =0.07 (ethyl acetate:hexane=3:1); FAB-MS: (M+H)⁺ =255.

EXAMPLE 36

With the exclusion of air, 172 mg (1.05 mmol) of 3-chloro-anilinehydrochloride (see Step 14.4) are added to 248 mg (0.70 mmol) ofN'-[3-(4-tert-butoxycarbonylamino-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidinein 2 ml of methanol and the reaction mixture is boiled for 19 hours. Thereaction mixture is then concentrated by evaporation and chromatographed(SiO₂, methylene chloride/ethanol [20:1]). Stirring with diethylether/hexane yields3-(4-tert-butoxycarbonylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;m.p. 165-168° C. (decomposition); FAB-MS: (M)⁺⁼ 436; HPLC: T_(Ret)(Grad₂₀₋₁₀₀)=15.4.

The starting material is prepared as follows:

Step 36.1:

Under an argon atmosphere, 254 mg (1.0 mmol) ofN'-[3-(4-amino-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidine(Step 35.4) in 4 ml of dioxane and 436.5 mg (2.0 mmol) of di-tert-butyldicarbonate are heated at 80° C. for 7 hours. Cooling, concentration byevaporation to a residual volume of ≈1 ml, the addition of ≈2 ml ofhexane and filtering yieldN'-[3-(4-tert-butoxycarbonylamino-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidine;m.p. 230-233° C. (decomposition); TLC: R_(f) =0.47 (methylenechloride:methanol=10:1); MS: (M)⁺ =354.

EXAMPLE 37

With the exclusion of air, 172.2 mg (1.05 mmol) of 3-chloro-anilinehydrochloride (see Step 14.4) are added to 199 mg (0.70 mmol) ofN'-[3-(3-nitro-phenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidinein 2 ml of methanol and the reaction mixture is boiled under reflux for19 hours. Cooling, filtering and washing with isopropanol and hexaneyield4-(3-chloro-phenylamino)-3-(3-nitro-phenyl)-1H-pyrazolo[3,4-d]pyrimidine;TLC: Rt=0.55 (methylene chloride:methanol=10:1); HPLC: T_(Ret)(Grad₂₀₋₁₀₀)=14.2; ¹ H-NMR: (DMSO-d₆) 9.05 (s, HN), 8.54 (s, 2H), 8.32(dd, J=8, 2, 1H), 8.22 (d, J=8, 1H), 7.8 (m, 2H), 7.49 (d, J=8, 1H),7.33 (t, J=8, 1H), 7.10 (dd, J=8, 2, 1H).

The starting material is prepared as follows:

Step 37.1:

Under a N₂ atmosphere, 13.47 g (0.42 mol) of sulfur and 60.72 g oftriethylamine are introduced into 65 ml of DMF. At 0-5° C., a solutionof 30 g (175 mmol) of 3-nitro-benzyl chloride in 35 ml of DMF is addeddropwise thereto and the reaction mixture is then stirred for 1.5 hoursat 0-5° C., for 3 hours at RT and finally for 4 hours at 40° C. (→exothermic). Then the orange reaction mixture is cooled to 0° C. and 13ml (208 mmol) of methyl iodide are added. The red suspension is stirredovernight at 0-5° C., poured into ice-water and then stirred for 1 hour.Ethyl acetate is added and the sulfur is filtered off. The aqueous phaseis separated off and extracted once with ethyl acetate. The organicphases are washed 3 times with water and brine, dried with Na₂ SO₄ andconcentrated by evaporation. Column chromatography (SiO₂, ethylacetate/hexane [1:4]) and crystallization from a diethyi ether solutionby the addition of hexane and cooling to -70° C. yield3-nitro-dithiobenzoic acid methyl ester; m.p. 38° C.

Step 37.2:

Under a N₂ atmosphere, 550 mg (3.8 mmol) of tetracyanoethylene oxide(Fluka, Buchs/Switzerland) are added to 500 mg (2.34 mmol) of3-nitro-dithiobenzoic acid methyl ester in 4 ml of toluene and thereaction mixture is then heated at 50° C. for 10 hours. 5 g of silicagel are added to the reaction mixture, which is then concentrated byevaporation. Application of the residue to a silica gel column, elutionwith hexane/ethyl acetate [1:2], treatment with activated carbon,concentration by evaporation and crystallization from diethyl ether(-70° C.) yield3-(3-nitrophenyl)-3-methylmercapto-2-cyano-acrylonitrile; MS: (M)⁺ =245,(M-SMe)⁺ =198, (M-NO₂ -Me)⁺ =184; IR: (KBr) 2222s, 1531s, 1514s, 1353s.

Step 37.3:

0.24 ml (4.8 mmol) of hydrazine hydrate is added dropwise to 1.00 g(4.08 mmol) of 3-(3-nitro-phenyl)-3-methylmercapto-2-cyano-acrylonitrilein 5.3 ml of methanol and the reaction mixture is then boiled for 1.5hours. The reaction mixture is then cooled in ice-water and theprecipitate is filtered off and washed with diethyl ether/isopropanol(2:1), yielding 5-amino-3-(3-nitro-phenyl)-1H-pyrazole-4-carbonitrile;TLC: R_(f) =0.4 (methylene chloride:methanol=10:1); T_(Ret)(Grad₂₀₋₁₀₀)=10.8; MS: (M)⁺ =229, (M-NO₂)⁺ =183.

Step 37.4:

In 8 ml of toluene, 229 mg (1.00 mmol) of5-amino-3-(3-nitro-phenyl)-1H-pyrazole-4-carbonitrile and 317 μl ofN,N-dimethylformamide dibenzylacetal are boiled overnight. Cooling,filtering the suspension and washing with hexane yieldN'-[3-(3-nitrophenyl)-4-cyano-1H-pyrazol-5-yl]-N,N-dimethylformamidine;m.p. 221-225° C.; T_(Ret) (Grad₂₂₋₁₀₀)=8.7.

EXAMPLE 38

In the presence of 30 mg of Raney nickel, 110 mg (0.30 mmol) of3-(3-nitro-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidineare hydrogenated in 3 ml of methanol and 3 ml of THF. The catalyst isfiltered off, then 15 g of silica gel are added to the filtrate, whichis then concentrated by evaporation. Application of the powder to asilica gel column, elution with methylene chloride/ethanol (15:1) andstirring of the crude product with diethyl ether/hexane yield3-(3-amino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo-[3,4-d]pyrimidine;m.p. 264-266° C.; TLC: R_(f) =0.45 (methylene chloride/methanol 10:1);HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=9.7.

EXAMPLE 39

The following compounds are obtained analogously to the processesdescribed in this text:

a)4-(3-chloro-phenylamino)-3-(4-[3-methyl-butanoylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,

b)4-(3-chloro-phenylamino)-3-(3-[3-methyl-butanoylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,

c)4-(3-chloro-phenylamino)-3-(4-propanoylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,

d)4-(3-chloro-phenylamino)-3-(3-propanoylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,

e)4-(3-chloro-phenylamino)-3-(4-pivaloylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine;HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=14.3, FAB-MS: (M+H)⁺ =421

f)4-(3-chloro-phenylamino)-3-(3-pivaloylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine;HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=15.06, FAB-MS: (M+H)⁺ =421,

g)3-(4-acetylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=11.4, FAB-MS: (M+H)⁺ =379,

h)3-(3-acetylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=12.0, FAB-MS: (M+H)⁺ =379,

i)4-(3-chloro-phenylamino)-3-(4-[{N,N-dimethylamino}-methyleneamino]-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,

j)4-(3-chloro-phenylamino)-3-(3-[{N,N-dimethylamino}-methyleneamino]-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,

k)4-(3-chloro-phenylamino)-3-(4-[thien-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,

l)4-(3-chloro-phenylamino)-3-(3-[thien-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=14,3, MS: (M)⁺ =421,

m)4-(3-chloro-phenylamino)-3-(4-[fur-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,

n)4-(3-chloro-phenylamino)-3-(3-[fur-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,

o)4-(3-chloro-phenylamino)-3-(4-[pyrid-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,

p)4-(3-chloro-phenylamino)-3-(3-[pyrid-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,

q)4-(3-chloro-phenylamino)-3-(4-methylsulfonylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,

r)4-(3-chloro-phenylamino)-3-(3-methylsulfonylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine;HPLC: T_(Ret) (Grad₂₀₋₁₀₀)=12,02, FAB-MS: (M+H)⁺ =415,

s)4-(3-chloro-phenylamino)-3-(4-[4-methyl-benzenesulfonylamino]-phenyl)-1H-pyrazolo-[3,4-d]pyrimidine,

t)4-(3-chloro-phenylamino)-3-(3-[4-methyl-benzenesulfonylamino]-phenyl)-1H-pyrazolo-[3,4-d]pyrimidine,

u)3-(3-tert-butoxycarbonylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,

v)3-(4-benzyloxycarbonylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidineand

w)3-(3-benzyloxycarbonylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine.

EXAMPLE 40

4-(3-Chloro-phenylamino)-3-(4-methoxy-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

A suspension of 1 g (3.52 mmol) of4-cyano-5-(dimethylamino-methyleneamino)-3-(4-methoxy-phenylamino)-pyrazoleand 0.9 g (5.48 mmol) of 3-chloro-aniline hydrochloride [for preparationsee: Step 14.4, cf. Justus Liebigs Ann. Chem. 176, 45 (1875)] in 7 ml ofmethanol is heated under reflux for 17 hours. The reaction mixture isthen cooled to room temperature, rendered alkaline (pH 10) by theaddition of 1N sodium hydroxide solution and filtered and the filterresidue is washed with a methanol/water mixture (1:1).

Recrystallization of the crude product from methanol/water yields thetitle compound having a water-content of 1.69% H₂ O; m.p. 223-224° C.(decomp.).

The starting material is prepared as follows:

Step 40.1:

4-Cyano-5-(dimethylamino-methyleneamino)-3-(4-methoxy-phenylamino)-pyrazole

A suspension of 5.41 g (23.6 mmol) of5-amino-4-cyano-3-(4-methoxy-phenylamino)-pyrazole [for preparation see:J. Heterocyclic Chem. 27, 775 (1990)] in 4.8 ml (27.2 mmol) ofN,N-dimethylformamide diethyl acetal (97%) and 100 ml of toluene isheated under reflux for 5 hours. The reaction mixture is then cooled toroom temperature and filtered and the filter residue is washed withtoluene. Recrystallization of the crude product from methanol/wateryields the title compound; m.p. 232-234° C. (decomp.).

EXAMPLE 41

3-(4-Acetylamino-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

A suspension of 6.32 g (20.3 mmol) of3-(4-acetylamino-phenylamino)-4-cyano-5-(dimethylamino-methyleneamino)-pyrazoleand 4 g (24.4 mmol) of 3-chloro-aniline hydrochloride in 50 ml ofmethanol is heated under reflux for 96 hours and then cooled to roomtemperature. The reaction mixture is filtered and the filter residue isdigested for 1/2 hour in 50 ml of 1N sodium hydroxide solution.Filtering and washing with water yield the title compound; m.p. 290-291°C. (decomp.).

The starting material is prepared as follows:

Step 41.1:

3-(4-Acetylamino-phenylamino)-2-cyano-3-methylmercapto-acrylonitrile

A mixture of 5 g (29.4 mmol) of3,3-bis-methylmercapto-2-cyano-acrylonitrile [for preparation see: Chem.Ber. 95, 2861 (1962)], 4.41 g (29.4 mmol) of 4-amino-acetanilide (Fluka)and 30 ml of toluene is heated under reflux for 20 hours. Cooling toroom temperature, filtering and washing the filter residue with tolueneyield the title compound; m.p 258-259° C. (decomp.).

Step 41.2:

3-(4-Acetylamino-phenylamino)-5-amino-4-cyano-pyrazole

A mixture of 7.46 g (27.4 mmol) of3-(4-acetylamino-phenylamino)-2-cyano-3-methylmercapto-acrylonitrile,1.63 ml (32.9 mmol) of hydrazine hydrate and 40 ml of methanol is heatedunder reflux for 5 hours and then concentrated by evaporation in vacuo.Recrystallization of the residue from methanol yields the titlecompound; m.p. 245-246° C. (decomp.).

Step 41.3:

3-(4-Acetylamino-phenylamino)-4-cyano-5-(dimethylamino-methyleneamino)-pyrazole

A suspension of 5.3 g (20.7 mmol) of3-(4-acetylamino-phenylamino)-5-amino-4-cyano-pyrazole in 4.4 ml (24.9mmol) of N,N-dimethylformamide diethyl acetal (97%) and 100 ml oftoluene is heated under reflux for 4 hours. The reaction mixture is thencooled to room temperature and filtered and the filter residue is washedwith toluene, yielding the title compound having a water content of4.22%; m.p. 275-276° C. (decomp.).

EXAMPLE 42

4-(3-Chloro-phenylamino)-3-(4-hydroxy-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

Under a nitrogen atmosphere, a solution of 5 ml (51.9 mmol) of borontribromide in 20 ml of methylene chloride is added dropwise over aperiod of 30 minutes to a suspension, cooled to 0° C., of 5 g (13.63mmol) of4-(3-chloro-phenylamino)-3-(4-methoxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidinein 100 ml of methylene chloride. The reaction mixture is stirred at 20°C. for 14 hours and then filtered and the filter residue is digested for1/2 hour at RT in 85 ml of water. After filtering, the filter residue ispartitioned between 50 ml of saturated sodium hydrogen carbonatesolution, 50 ml of water and 135 ml of THF. The organic phase isseparated off, the aqueous phase is extracted again with 35 ml of THF,the combined THF extracts are washed with brine and the organic phase isdried over sodium sulfate and concentration by evaporation is carriedout in vacuo. The crystalline residue is digested in 65 ml of boilingethyl acetate, cooled to RT and filtered, yielding the crude titlecompound. Recrystallization of a sample from ethyl acetate yields thepure title compound; m.p. >260° C.; TLC-R_(f) =0.54(toluene/isopropanol/conc. ammonia [70:29:1]).

EXAMPLE 43

4-(3-Chloro-phenylamino)-3-(4-dimethylamino-phenylamino)-1H-pyrazolo-[3,4-d]pyrimidine

A mixture of 5 g (16.8 mmol) of4-cyano-5-(dimethylamino-methyleneamino)-3-(4-dimethylamino-phenylamino)-pyrazole,3.3 g (20.1 mmol) of 3-chloro-aniline hydrochloride and 40 ml of DMF isheated for 8 hours, with stirring, at 130° C. The reaction mixture isthen cooled to RT, water is added dropwise thereto, and the reactionmixture is then filtered and the filter residue is dissolved in approx.20 ml of DMF. After precipitation with water, followed byrecrystallization from DMF/water, the resulting crystals are suspendedin 10 ml of water. 6 ml of 1N hydrochloric acid are added, the mixtureis heated briefly to reflux and filtered and 3 ml of 2N sodium hydroxidesolution are added to the filtrate. The crystalline precipitate isfiltered off, washed with water and dried under a high vacuum at 120°C., yielding the title compound having a water content of 2.32%; m.p.250-255° C. (decomp.).

The starting material is prepared as follows:

Step 43.1:

4-Cyano-5-(dimethylamino-methyleneamino)-3-(4-dimethylamino-phenylamino)-pyrazole

A suspension of 10 g (41.3 mmol) of5-amino-4-cyano-3-(4-dimethylamino-phenylamino)-pyrazole [forpreparation see: Arch.Pharm.(Weinheim) 326, 245 (1993)] in 8.75 ml (49.5mmol) of N,N-dimethylformamide diethyl acetal (97%) and 200 ml oftoluene is heated under reflux for 1 hour. The reaction mixture is thencooled to approx. 30° C. and filtered and the filter residue is washedwith toluene, yielding the title compound; m.p. 281-282° C. (decomp.).

EXAMPLE 44

4-(3-Chloro-phenylamino)-3-(4-methoxy-benzylamino)-1H-pyrazolo[3,4-d]-pyrimidine

A mixture of 4.475 g (15 mmol) of4-cyano-5-(dimethylamino-methyleneamino)-3-(4-methoxy-benzylamino)-pyrazole,2.83 g (17.25 mmol) of 3-chloro-aniline hydrochloride and 80 ml ofethanol is heated under reflux for 16 hours. Cooling to approx. 5° C.,filtering and washing the filter residue with diethyl ether yield thetitle compound; m.p. 222-223° C.

The starting material is prepared as follows:

Step 44.1:

2-Cyano-3-(4-methoxy-benzylamino)-3-methylmercapto-acrylonitrile

A mixture of 17.03 g (0.1 mol) of3,3-bis-methylmercapto-2-cyano-acrylonitrile, 12.98 ml (0.1 mol) of4-methoxy-benzylamine and 60 ml of ethanol is heated under reflux for 2hours. Approx. 90 ml of hexane are then added dropwise to the hotsolution which is then allowed to cool to RT. The title compound whichhas precipitated in the form of colorless crystals is filtered off,washed with diethyl ether and dried under a HV; m.p. 100-101° C.

Step 44.2:

5-Amino-4-cyano-3-(4-methoxy-benzylamino)-pyrazole

A mixture of 15 g (57.8 mmol) of2-cyano-3-(4-methoxy-benzylamino)-3-methylmercapto-acrylonitrile, 3.03ml (61.1 mmol) of hydrazine hydrate and 40 ml of methanol is stirred for1 hour at RT, heated under reflux for 2 hours and then concentrated byevaporation in vacuo. Recystallization of the residue fromethanol/hexane yields the title compound; m.p. 148-149° C.

Step 44.3:

4-Cyano-5-(dimethylamino-methyleneamino)-3-(4-methoxy-benzylamino)-pyrazole

A mixture of 6.08 g (25 mmol) of5-amino-4-cyano-3-(4-methoxy-benzylamino)-pyrazole, 5.14 ml (30 mmol) ofN,N-dimethylformamide diethyl acetal and 90 ml of toluene is heatedunder reflux for 3 hours. The reaction mixture is then cooled to approx.5° C. and filtered and the filter residue is washed with toluene,yielding the title compound; m.p. 147-148° C.

EXAMPLE 45

4-(3-Chloro-phenylamino)-3-(3-methoxy-benzylamino)-1H-pyrazolo[3,4-d]-pyrimidine(title compound I) and5-(3-chloro-phenyl)-1,5-dihydro-4-imino-3-(3-methoxy-benzylamino)-4H-pyrazolo[3,4-d]pyrmidinehydrochloride (title compound II)

A mixture of 5.97 g (20 mmol) of4-cyano-5-(dimethylamino-methyleneamino)-3-(3-methoxy-benzylamino)-pyrazole,3.77 g (23 mmol) of 3-chloro-aniline hydrochloride and 100 ml of ethanolis heated under reflux for 36 hours. Cooling to approx. 10° C.,filtering and washing the filter residue with ethanol yield titlecompound II; m.p. 251-253° C. (decomp.). The filtrate is concentrated byevaporation in vacuo, the oily residue is partitioned between ethylacetate and water and the organic phase is washed with brine, dried oversodium sulfate and concentrated to a volume of approx. 25 ml using anRV, a crystalline precipitate being formed. Filtering and washing thefilter residue with a small amount of ethyl acetate and diethyl etheryield title compound I (see Example 52); m.p. 194-195° C.

The starting material is prepared as follows:

Step 45.1:

2-Cyano-3-(3-methoxy-benzylamino)-3-methylmercapto-acrylonitrile

A mixture of 17.03 g (0.1 mol) of3,3-bis-methylmercapto-2-cyano-acrylonitrile, 12.78 ml (0.1 mol) of3-methoxy-benzylamine and 60 ml of ethyl acetate is heated under refluxfor 2 hours and then concentrated by evaporation in vacuo, yielding theoily title compound; TLC-R_(f) 32 0.28 (toluene/isopropanol [9:1]).

Step 45.2:

5-Amino-4-cyano-3-(3-methoxy-benzylamino)-pyrazole

A mixture of 25.57 g (98.6 mmol) of2-cyano-3-(3-methoxy-benzylamino)-3-methylmercapto-acrylonitrile, 5.16ml (104 mmol) of hydrazine hydrate and 140 ml of methanol is stirred atRT for 1 hour, heated under reflux for 2.5 hours and then concentratedby evaporation in vacuo. Recrystallization of the residue fromethanol/hexane yields the title compound; m.p. 151-153° C.

Step 45.3:

4-Cyano-5-(dimethylamino-methyleneamino)-3-(3-methoxy-benzylamino-pyrazole

A mixture of 9.12 g (37.5 mmol) of5-amino-4-cyano-3-(3-methoxy-benzylamino)-pyrazole, 7.71 ml (45 mmol) ofN,N-dimethylformamide diethyl acetal and 130 ml of toluene is heatedunder reflux for 3 hours and then approx. 40 ml of hexane are addeddropwise thereto. Cooling to 10° C. causes a crystalline precipitate toform. Filtering and washing the filter residue with diethyl ether yieldthe title compound; m.p. 136-137° C.

EXAMPLE 46

4-Benzylamino-3-(3-methyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

Under a nitrogen atmosphere, a mixture of 1 g (3.74 mmol) of4-cyano-5-(dimethylamino-methyleneamino)-3-(3-methyl-phenylamino)-pyrazoleand 10 ml of benzylamine is stirred for 4 hours at 120° C. and thenconcentrated by evaporation under a HV. The crystalline residue isdigested in 10 ml of acetonitrile and filtered and the filter residue isrecrystallized from 40 ml of acetonitrile, yielding the title compound;m.p. 200-201° C.

The starting material is prepared as follows:

Step 46.1:

4-Cyano-5-(dimethylamino-methyleneamino)-3-(3-methyl-phenylamino)-pyrazole

A suspension of 10.71 g (50.22 mmol) of5-amino-4-cyano-3-(3-methyl-phenylamino)-pyrazole [for preparation see:Arch. Pharm. (Weinheim) 326, 245 (1993)], 9.9 ml (57.8 mmol) ofN,N-dimethylformamide diethyl acetal and 150 ml of toluene is heatedunder reflux for 4 hours. The reaction mixture is then cooled to RT andfiltered and the filter residue is washed with toluene and diethylether, yielding the title compound; m.p. 260-261° C.

EXAMPLE 47

(S)-3-(3-Methyl-phenylamino)-4-(1-phenyl-ethylamino)-1H-pyrazolo[3,4-d]-pyrimidinehydrochloride

Under a nitrogen atmosphere, a mixture of 1 g (3.74 mmol) of4-cyano-5-(dimethylamino-methyleneamino)-3-(3-methyl-phenylamino)-pyrazoleand 10 ml of (S)-1-phenyl-ethylamine is stirred at 120° C. for 24 hoursand then concentrated by evaporation under a HV. The oily residue ispurified by flash chromatography on silica gel having a particle size of0.04-0.06 mm using ethyl acetate/hexane (7:3). The product-containingfractions are concentrated by evaporation and the resinous residue isdissolved in a mixture of 5 ml of ethanol and 0.8 ml of 4N hydrochloricacid. Concentration by evaporation again and recrystallization of theresidue from ethanol/diethyl ether yield the title compound; m.p.170-180° C. (decomp.); [a]_(D) ²⁰ =+253.8±1° (c=1.015%, methanol).

EXAMPLE 48

(R)-3-(3-Methyl-phenylamino)-4-(1-phenyl-ethylamino)-1H-pyrazolo[3,4-d]-pyrimidine hydrochloride

Analogously to Example 47, the title compound [m.p. 170-180° C.(decomp.); [a]_(D) ²⁰ =-249.4±1° (c=0.987%, methanol)] is obtained from1.5 g (5.61 mmol) of4-cyano-5-(dimethylamino-methyleneamino)-3-(3-methyl-phenylamino)-pyrazoleand 10 ml of (R)-1-phenyl-ethylamine.

EXAMPLE 49

4-Benzylamino-3-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

Under a nitrogen atmosphere, a mixture of 1 g (3.46 mmol) of3-(3-chloro-phenylamino)-4-cyano-5-(dimethylamino-methyleneamino)-pyrazoleand 10 ml of benzylamine is stirred for 3 hours at 120° C. and thenconcentrated by evaporation under a HV. The crystalline residue isdigested in 10 ml of acetonitrile, cooled to 0° C. and filtered and thefilter residue is recrystallized from 35 ml of acetonitrile, yieldingthe title compound; m.p. 216-218° C.

The starting material is prepared as follows:

Step 49.1:

5-Amino-3-(3-chloro-phenylamino)-4-cyano-pyrazole

A mixture of 15 g (60.1 mmol) of3-(3-chloro-phenylamino)-2-cyano-3-methylmercapto-acrylonitrile [forpreparation see: EP 0 010 396 A1], 3.13 ml (63.2 mmol) of hydrazinehydrate and 100 ml of methanol is heated under reflux for 3 hours andthen concentrated by evaporation in vacuo. Recrystallization of theresidue from ethyl acetate/hexane yields the title compound; m.p.195-200° C.

Step 49.2:

3-(3-Chloro-phenylamino)-4-cyano-5-(dimethylamino-methyleneamino)-pyrazole

Analogously to Step 44.3, the title compound [m.p. >260° C.; TLC-R_(f)=0.87 (methylene chloride/methanol [9:1])] is obtained from 11.7 g(50.07 mmol) of 5-amino-3-(3-chloro-phenylamino)-4-cyano-pyrazole, 9.43ml (55.07 mmol) of N,N-dimethylformamide diethyl acetal and 140 ml oftoluene after heating under reflux for 3.5 hours.

EXAMPLE 50

4-(3-Chloro-phenylamino)-3-(3-methoxy-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

A mixture of 1.38 g (4.85 mmol) of4-cyano-5-(dimethylamino-methyleneamino)-3-(3-methoxy-phenylamino)-pyrazole,0.915 g (5.58 mmol) of 3-chloro-aniline hydrochloride and 20 ml ofmethanol is heated under reflux for 16 hours. Concentration of thereaction mixture by evaporation in vacuo and recrystallization of theresidue from methanol/water yield the title compound; m.p. 202-203° C.

The starting material is prepared as follows:

Step 50.1:

5-Amino-4-cyano-3-(3-methoxy-phenylamino)-pyrazole

Analogously to Example 49.1, the title compound [m.p. 177-180° C.] isobtained from 10 g (40.77 mmol) of2-cyano-3-(3-methoxy-phenylamino)-3-methylmercapto-acrylonitrile [forpreparation see: Bioorg. Med. Chem. Lett. 4, 615 (1994)], 2.12 ml (42.8mmol) of hydrazine hydrate and 70 ml of methanol after heating underreflux for 3.5 hours.

Step 50.2:

4-Cyano-5-(dimethylamino-methyleneamino)-3-(3-methoxy-phenylamino)-pyrazole

Analogously to Example 44.3, the title compound [m.p. 227-231° C.] isobtained from 1.25 g (5.45 mmol) of5-amino-4-cyano-3-(3-methoxy-phenylamino)-pyrazole, 1.23 ml (7.18 mmol)of N,N-dimethylformamide diethyl acetal and 25 ml of toluene afterheating under reflux for 4.5 hours.

EXAMPLE 51

4-(3-Chloro-phenylamino)-3-(3-methoxy-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

A mixture of 0.9 g (2.45 mmol) of5-(3-chloro-phenyl)-1,5-dihydro-4-imino-3-(3-methoxy-phenylamino)-4H-pyrazolo[3,4-d]pyrimidine,18 ml of dioxane and 18 ml of water is heated under reflux for 10 hours.Cooling to 0° C., filtering and recrystallization from methanol/waterand from methanol yield the title compound; m.p. 203-204° C.

The starting material is prepared as follows:

Step 51.1:

4-Cyano-5-(ethoxy-methyleneamino)-3-(3-methoxy-phenylamino)-pyrazole

With stirring, a mixture of 4 g (17.45 mmol) of5-amino-4-cyano-3-(3-methoxy-phenylamino)-pyrazole (Step 50.1) and 50 mlof orthoformic acid triethyl ester is heated at 120° C. for 1.5 hours,care being taken to ensure that the ethanol formed in the course of thereaction is distilled off from the reaction mixture. The reactionmixture is cooled to 30° C., approx. 100 ml of diethyl ether are addedand the reaction mixture is cooled further to 0° C. and filtered.Washing with diethyl ether yields the title compound; m.p. 181-183° C.

Step 51.2:

5-(3-Chloro-phenyl)-1.5-dihydro-4-imino-3-(3-methoxy-phenylamino)-4H-pyrazolo[3,4-d]pyrimidine

A mixture of 3.85 g (13.49 mmol) of4-cyano-5-(ethoxy-methyleneamino)-3-(3-methoxy-phenylamino)-pyrazole,2.84 ml (27.02 mmol) of 3-chloro-aniline and 75 ml of ethanol is heatedunder reflux for 2 hours. The reaction mixture is then cooled to RT andfiltered. Washing the crystals with ethanol and diethyl ether yields thetitle compound; m.p. 190-192° C.

EXAMPLE 52

4-(3-Chloro-phenylamino)-3-(3-methoxy-benzylamino)-1H-pyrazolo[3,4-d]-pyrimidine

A mixture of 0.417 g (1 mmol) of5-(3-chloro-phenyl)-1,5-dihydro-4-imino-3-(3-methoxy-benzylamino)-4H-pyrazolo[3,4-d]pyrimidinehydrochloride (Example 45, title compound II), 20 ml of dioxane, 19 mlof water and 1 ml of 1N sodium hydroxide solution is heated under refluxfor 10 hours. Cooling to approx. 5° C., filtering, washing the filterresidue with water and drying under a HV at 120° C. yield the titlecompound (see Example 45); m.p. 192-193° C.

EXAMPLE 53

3-(3-Hydroxy-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

With the exclusion of air and moisture, a mixture of 44.2 mg (0.1 mmol)of1-benzyl-4-(3-chloro-phenylamino)-3-(3-methoxy-phenyl)-pyrazolo[3,4-d]pyrimidine,80 mg (0.6 mmol) of anhydrous aluminum chloride and 2 ml of toluene isheated under reflux for 2 hours. The reaction mixture is then pouredinto ice-water and extracted with ethyl acetate. The organic phase iswashed with a saturated solution of sodium hydrogen carbonate in waterand with brine, dried over sodium sulfate and concentrated byevaporation in vacuo. Purification of the residue by flashchromatography on silica gel (230-400 mesh) using methylene chloride andmethylene chloride/methanol mixtures (19:1 and 17:3, respectively)yields the title compound; m.p. >220° C.; FAB-MS: (M+H)⁺ =338.

The starting material is prepared as follows:

Step 53.1:

2-Cyano-3-(3-methoxy-phenyl)-3-methylmercapto-acrylonitrile

A mixture of 4.88 g (24.61 mmol) of 3-methoxy-dithiobenzoic acid methylester [for preparation see: Chem. Ber. 120,1757 (1987)], 4.26 g (29.56mmol) of tetracyanoethylene oxide (Fluka) and 35 ml of toluene isstirred for 1 hour at -5° C. and for 21 hours at 20° C. Then 2 ml oftriethylamine are added to the reaction mixture, which is thenconcentrated while passing a stream of nitrogen through the reactionmixture for approx. 15 hours. The oily residue is purified by flashchromatography on silica gel (230-400 mesh) using hexane andhexane/-ethyl acetate mixtures having an increasing content of ethylacetate (5%, 7.5%, 10%, 12.5% and 20% ethyl acetate). Theproduct-containing fractions are concentrated by evaporation in vacuo,yielding the title compound in the form of an oil; TLC-R_(f) =0.48(toluene/acetone [9:1]); FAB-MS: (M+H)⁺ =231 (C₁₂ H₁₀ N₂ OS).

Step 53.2:

5-Amino-1-benzyl-4-cyano-3-(3-methoxy-phenyl)-pyrazole

At 5° C., 11.94 ml of a 5.4N solution of sodium methanolate in methanol(64.47 mmol) is added to a suspension of 6 g (30.62 mmol) ofbenzylhydrazine dihydrochloride in 15 ml of ethanol and the reactionmixture is stirred for approx. 15 minutes at 5-10° C. and thenintroduced into a suspension of 2.95 g (12.81 mmol) of2-cyano-3-(3-methoxy-phenyl)-3-methyl-mercapto-acrylonitrile in 95 ml ofethanol. The reaction mixture is heated under reflux for 1 hour, thencooled to RT and filtered. The filter residue is washed with ethanol andthe filtrate is concentrated by evaporation in vacuo. The oily residueis purified by flash chromatography on silica gel (230-400 mesh) using atoluene/acetone mixture (19:1). The combined product-containingfractions are concentrated by evaporation and the residue is digested indiisopropyl ether. Filtration yields the title compound; m.p. 147-149°C.; FAB-MS: (M+H)⁺ =305.

Step 53.3:

1-Benzyl-4-hydroxy-3-(3-methoxy-phenyl)-pyrazolo[3,4-d]pyrimidine

A mixture of 2.55 g (8.38 mmol) of5-amino-1-benzyl-4-cyano-3-(3-methoxy-phenyl)-pyrazole and 20 ml of 85%aqueous formic acid is heated under reflux for 21 hours and then cooledto room temperature and, with stirring, a small amount of water is addedthereto. Filtering and washing the filter residue with water yield thetitle compound; m.p. 183-185° C.; FAB-MS: (M+H)⁺ =333.

Step 53.4:

1-Benzyl-4-chloro-3-(3-methoxy-phenyl)-pyrazolo[3,4-d]pyrimidine

A mixture of 2.48 g (7.46 mmol) of1-benzyl-4-hydroxy-3-(3-methoxy-phenyl)-pyrazolo[3,4-d]pyrimidine and 30ml of phosphorus oxychloride is heated under reflux for 10 hours andthen, with stirring, poured slowly into ice-water. The reaction mixtureis stirred for a further 2 hours at 0-10° C. and filtered and the filterresidue is washed with water. The crude product is dissolved inmethylene chloride and the organic phase is washed with brine, driedover sodium sulfate and concentrated by evaporation in vacuo.Purification of the residue by flash chromatography on silica gel(230-400 mesh) using methylene chloride and methylene chloride/methanolmixtures (99:1 and 49:1) yields the title compound; m.p. 99-101° C.;FAB-MS: (M+H)⁺ =351.

Step 53.5:

1-Benzyl-4-(3-chloro-phenylamino)-3-(3-methoxy-phenyl-pyrazolo[3,4-d]-pyrimidine

A mixture of 70 mg (0.2 mmol) of1-benzyl-4-chloro-3-(3-methoxy-phenyl)-pyrazolo[3,4-d]-pyrimidine, 115.6μl (1.1 mmol) of 3-chloro-aniline and 7 ml of 1-butanol is heated underreflux for 20 hours and then concentrated by evaporation in vacuo. Theresidue is digested in hexane and filtered. Repeated digestion of thefilter residue in diisopropyl ether and in ethanol yields the titlecompound; m.p. 118-119° C.

EXAMPLE 54

4-(3-Chloro-phenylamino)-3-(4-hydroxy-phenyl)-1H-pyrazolo[3,4-d]pyrimidine

With the exclusion of air and moisture, a mixture of 0.442 g (1.2 mmol)of4-(3-chloro-phenylamino)-3-(4-methoxy-phenyl)-1H-pyrazolo[3,4-d]pyrimidine[see Example 34], 1.16 g (8.7 mmol) of anhydrous aluminum chloride and20 ml of benzene is heated under reflux for 45 minutes. The reactionmixture is then poured into ice-water, filtered and washed with waterand the filter residue is partitioned between a 5% solution of sodiumhydrogen carbonate in water and ethyl acetate. The organic phase iswashed with brine, dried over sodium sulfate and concentrated in vacuo.A small amount of hexane is added and the desired product precipitates.Washing with hexane and purification of the crude product by flashchromatography on silica gel (230-400 mesh) using an ethylacetate/hexane mixture (7:3) yield the title compound; m.p. >220° C.;MS: (M)⁺ =337.

EXAMPLE 55

(R)-3-(3-Chloro-phenylamino)-4-(1-phenyl-ethylamino)-1H-pyrazolo[3,4-d]-pyrimidinehydrochloride

Under a nitrogen atmosphere, a mixture of 1.5 g (5.19 mmol) of3-(3-chloro-phenylamino)-4-cyano-5-(dimethylamino-methyleneamino)-pyrazole(Step 49.2) and 10 ml of (R)-1-phenylethylamine is stirred at 120° C.for 40 hours and then concentrated by evaporation under a HV. The oilyresidue is purified by flash chromatography on silica gel having aparticle size of 0.04-0.06 mm using ethyl acetate/hexane (7:3). Theproduct-containing fractions are concentrated by evaporation and theresinous residue is dissolved in a mixture of 5 ml of ethanol and 1.4 mlof 4N hydrochloric acid. Concentration by evaporation again andrecrystallization of the residue from ethanol/diethyl ether and fromethanol yield the title compound; m.p. 150-152° C.; EI-MS: (M)⁺ =364;[a]_(D) ²⁰ =-233.7±2.1° (c=0.486% methanol).

EXAMPLE 56

(S)-3-(3-Chloro-phenylamino)-4-(1-phenyl-ethylamino)-1H-pyrazolo[3,4-d]-pyrimidinehydrochloride

0.4 g (1.1 mmol) of(S)-3-(3-chloro-phenylamino)-1,5-dihydro-4-imino-5-(1-phenyl-ethyl)-4H-pyrazolo[3,4-d]pyrimidinein the molten state is stirred for 15 minutes at 220° C. The crudeproduct obtained after cooling to RT is purified by flash chromatographyon silica gel having a particle size of 0.04-0.06 mm using ethylacetate/hexane (7:3). The product-containing fractions are concentratedby evaporation and the resinous residue is dissolved in ethanol. Theaddition of 0.6 ml of 4N hydrochloric acid, concentration by evaporationin vacuo and recrystallization of the residue from ethanol yield thetitle compound; m.p. 153-155° C.; EI-MS: (M)⁺ =364; [a]_(D) ²⁰=+232.7±2.2° (c=0.465%, methanol).

The starting material is prepared as follows:

Step 56.1:

3-(3-Chloro-phenylamino)-4-cyano-5-(ethoxy-methyleneamino)-pyrazole

Analogously to Step 51.1, the title compound [m.p. 197-199° C.] isobtained from 4.5 g (19.26 mmol) of5-amino-3-(3-chloro-phenylamino)-4-cyano-pyrazole (Step 49.1) and 50 mlof orthoformic acid triethyl ester after stirring for 2 hours at 120° C.

Step 56.2:

(S)-1,5-Dihydro-3-(3-chloro-phenylamino)-4-imino-5-(1-phenyl-ethyl)-4H-pyrazolo[3,4-d]pyrimidine

Analogously to Step 51.2, the title compound [m.p. 214-215° C.; [a]_(D)²⁰ =-279.6±1° (c=0.98%, methanol)] is obtained from 1.2 g (4.14 mmol) of3-(3-chloro-phenylamino)-4-cyano-5-(ethoxy-methyleneamino)-pyrazole,0.632 ml (4.97 mmol) of (S)-1-phenyl-ethylamine and 12 ml of ethanolafter heating under reflux for 2 hours.

EXAMPLE 57

3-(4-Acetylamino-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

A mixture of 0.34 g (1.04 mmol) of3-(4-acetylamino-benzylamino)-4-cyano-5-(dimethylamino-methyleneamino)-pyrazole,0.189 g (1.15 mmol) of 3-chloro-aniline hydrochloride and 4 ml ofmethanol is heated under reflux for 20 hours. Cooling to approx. 20° C.,filtering and washing the filter residue with diethyl ether yields thetitle compound; m.p. >260° C.; TLC-R_(f) =0.34(toluene/isopropanol/conc. ammonia [70:29:1]).

The starting material is prepared as follows:

Step 57.1:

3-(4-Amino-benzylamino)-2-cyano-3-methylmercapto-acrylonitrile

A mixture of 17.03 g (0.1 mol) of3,3-bis-methylmercapto-2-cyano-acrylonitrile, 11.34 ml (0.1 mol) of4-amino-benzylamine (Aldrich) and 60 ml of ethanol is stirred at 50° C.for 2 hours and then concentrated by evaporation in vacuo. Purificationof the residue by flash chromatography on silica gel having a particlesize of 0.04-0.06 mm using toluene/isopropanol mixtures (49:1, 97:3,24:1 and 9:1) yields the title compound; m.p. 100-102° C.

Step 57.2:

5-Amino-3-(4-amino-benzylamino)-4-cyano-pyrazole

A mixture of 15.71 g (64.3 mmol) of3-(4-amino-benzylamino)-2-cyano-3-methylmercapto-acrylonitrile, 3.35 ml(67.5 mmol) of hydrazine hydrate and 90 ml of methanol is stirred for 1hour at 20° C., heated under reflux for 4 hours and then concentrated byevaporation in vacuo. Digestion of the crystalline residue in 70 ml ofdiisopropyl ether, filtering and digestion again in 100 ml ofisopropanol yield the title compound; m.p. 168-170° C.

Step 57.3:

3-(4-Acetylamino-benzylamino)-5-amino-4-cyano-pyrazole

With stirring, a solution of 1.087 ml (11.5 mmol) of acetic anhydride in20 ml of THF is added dropwise over a period of 15 minutes, to asuspension, cooled to 0° C., of 2.5 g (10.95 mmol) of5-amino-3-(4-amino-benzylamino)-4-cyano-pyrazole in 50 ml of THF. Thereaction mixture is stirred for a further 2 hours at RT, there initiallybeing formed a solution from which crystalline product graduallyprecipitates. The reaction mixture is filtered and the filter residue iswashed with THF and diethyl ether. Drying under a HV (8 hours, 110° C.)yields the title compound containing 21.06% THF; m.p. 124-126° C.(decomp.); FAB-MS: (M+H)⁺ 271 (C₁₃ H₁₄ N₆ O).

Step 57.4:

3-(4-Acetylamino-benzylamino)-4-cyano-5-(dimethylamino-methyleneamino)-pyrazole

A mixture of 0.48 g (1.4 mmol) of3-(4-acetylamino-benzylamino)-5-amino-4-cyano-pyrazole containing 21.06%THF, 0.245 ml (1.43 mmol) of N,N-dimethylformamide diethyl acetal and 4ml of toluene is heated under reflux for 4.5 hours. The reaction mixtureis then cooled to approx. 0° C. and filtered and the filter residue iswashed with toluene, yielding the title compound; m.p. 206-211° C.

EXAMPLE 58

3-(4-Amino-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidinehydrochloride

A mixture of 0.25 g (0.553 mmol) of3-[4-(N-BOC-amino)-phenylamino]-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine(see Example 65) and 10 ml of a 3N solution of hydrochloric acid inmethanol is stirred at RT for 20 hours. Then 10 ml of diethyl ether areadded, the reaction mixture is filtered and the filter residue isdigested in hot methanol. After cooling, filtering and washing thefilter residue with diethyl ether, the crystals are dried for 15 hoursunder a high vacuum at 100° C. and then left to stand under ambientconditions for 24 hours, yielding the title compound containing 10.97%hydrogen chloride and 4.34% water; m.p. 211-213° C.; EI-MS: (M)⁺ =351.

EXAMPLE 59

4-(3-Chloro-phenylamino)-3-(3-hydroxy-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

With the exclusion of air and moisture, a mixture of 0.5 g (1.36 mmol)of4-(3-chloro-phenylamino)-3-(3-methoxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine(Example 50), 0.873 g (6.55 mmol) of anhydrous aluminum chloride and 15ml of benzene is heated at 80° C. for 9 hours. The benzene phase is thendecanted off, the resinous residue is partitioned between ethyl acetateand water and the organic phase is washed with water and with asaturated solution of sodium hydrogen carbonate in water, dried oversodium sulfate and concentrated by evaporation in vacuo. The residue ispurified by flash chromatography on silica gel having a particle size of0.04-0.06 mm using methylene chloride/methanol mixtures (50:1 and 20:1).The product-containing fractions are combined and concentrated to avolume of approx. 10 ml, the desired product crystallising out.Filtering and washing the filter residue with diethyl ether yield thetitle compound; m.p. 265-266° C.; EI-MS: (M)⁺ =352.

EXAMPLE 60

4-Benzylamino-3-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

A mixture of 1 g (4.28 mmol) of5-amino-3-(3-chloro-phenylamino)-4-cyano-pyrazole (Step 49.1), 3.67 g(34.25 mmol) of benzylamine, 0.245 ml (4.28 mmol) of glacial acetic acidand 0.73 ml (19.35 mmol) of formic acid is heated at 200° C. for 20hours. The reaction mixture is cooled to RT, 30 ml of ice-water and 5 mlof ethanol are added and stirring is carried out for a further 20minutes. The reaction mixture is then filtered and the filter residue iswashed with water. Recrystallization from isopropyl alcohol yields thetitle compound; m.p. 215-218° C.

EXAMPLE 61

4-Benzylamino-3-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

A mixture of 1 g (4.28 mmol) of5-amino-3-(3-chloro-phenylamino)-4-cyano-pyrazole (Step 49.1) and 1.75 g(12.95 mmol) of N-benzylformamide (Aldrich) is heated at 200° C. for 20hours. The reaction mixture is purified by flash chromatography onsilica gel having a particle size of 0.04-0.06 mm using ethyl acetate.Concentration of the product-containing fractions by evaporation andrecrystallization of the residue from ethyl acetate/hexane yield thetitle compound; m.p. 215-218° C.; FAB-MS: (M+H)⁺ =351.

EXAMPLE 62

3-(4-Amino-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

A mixture of 0.11 g (0.27 mmol) of3-(4-acetylamino-benzylamino)-4-(3-chlorophenylamino)-1H-pyrazolo[3,4-d]pyrimidine(Example 57), 3 ml of water and 1.5 ml of 30% sodium hydroxide solutionis heated at 100° C. for 4 hours. After cooling to RT, the reactionmixture is extracted with ethyl acetate and the extract is washed withbrine, dried over sodium sulfate and concentrated by evaporation invacuo. The residue is purified by flash chromatography on silica gelhaving a particle size of 0.04-0.06 mm using methylene chloride/methanolmixtures (50:1 and 20:1). The product-containing fractions areconcentrated by evaporation and the residue is recrystallized from ethylacetate/hexane, yielding the title compound; m.p. 170-171° C.; EI-MS:(M)⁺ =365.

EXAMPLE 63

4-(3-Chloro-phenylamino)-3-(4-ethoxy-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

A mixture of 5.8 g (19.44 mmol) of4-cyano-5-(dimethylamino-methyleneamino)-3-(4-ethoxy-phenylamino)-pyrazole,3.51 g (21.4 mmol) of 3-chloro-aniline hydrochloride and 40 ml ofmethanol is heated under reflux for 15 hours. Cooling to 10° C.,filtering and washing the filter residue with methanol and diethyl etheryield the title compound; m.p. 232-233° C.; EI-MS: (M)⁺ =380.

The starting material is prepared as follows:

Step 63.1:

2-Cyano-3-(4-ethoxy-phenylamino)-3-methylmercapto-acrylonitrile

A mixture of 17.03 g (0.1 mol) of3,3-bis-methylmercapto-2-cyano-acrylonitrile, 12.94 ml (0.1 mol) of4-ethoxy-aniline and 60 ml of ethanol is heated under reflux for 2hours. Then, with stirring, 120 ml of diethyl ether are added dropwiseto the reaction mixture, which is at approx. 30° C. The reaction mixtureis then cooled to 0° C. The title compound precipitates in the form ofcolorless crystals and is filtered off, washed with diethyl ether anddried under a HV; m.p. 141-142° C.

Step 63.2:

5-Amino-4-cyano-3-(4-ethoxy-phenylamino)-pyrazole

A mixture of 21.5 g (82.9 mmol) of2-cyano-3-(4-ethoxy-phenylamino)-3-methylmercapto-acrylonitrile, 4.31 ml(87 mmol) of hydrazine hydrate and 110 ml of methanol is heated underreflux for 7 hours and then concentrated by evaporation in vacuo.Recrystallization of the residue from ethyl acetate/hexane yields thetitle compound; m.p. 166-167° C.

Step 63.3:

4-Cyano-5-(dimethylamino-methyleneamino)-3-(4-ethoxy-phenylamino)-pyrazole

A suspension of 4.86 g (19.98 mmol) of5-amino-4-cyano-3-(4-ethoxy-phenylamino)-pyrazole in 3.94 ml (23 mmol)of N,N-dimethylformamide diethyl acetal and 60 ml of toluene is heatedunder reflux for 2 hours. The reaction mixture is then cooled to 20° C.and filtered and the filter residue is washed with toluene, yielding thetitle compound; m.p. 246-247° C. (decomp.).

EXAMPLE 64

The following compounds are obtained analogously to the processesdescribed in this text, e.g. in Examples 40-63:

a)4-(3-chloro-phenylamino)-3-(3,4-dimethoxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;m.p. 168-170° C.

b)4-(3-chloro-phenylamino)-3-(3,5-dimethoxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;m.p. 21 8-220cC

c)4-(3-chloro-phenylamino)-3-(3-formylamino-4-methoxy-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

d)3-(3-acetylamino-4-methoxy-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

e)3-(4-acetylamino-3-methoxy-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine

f)4-(3-chloro-phenylamino)-3-(4-formylamino-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

g)4-(3-chloro-phenylamino)-3-(4-formylamino-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine

h)4-(3-chloro-phenylamino)-3-(4-propionylamino-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine

i)3-(4-aminomethyl-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidinehydrate; m.p. 207-209° C.

k)3-(3-aminomethyl-phenylainino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

l)3-[4-(N-BOC-aminomethyl)-phenylamino]-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine-hydrate[precursor of i)]; m.p. 196° C.

m)3-[3-(N-BOC-aminomethyl)-phenylamino]-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine[precursor of k)].

n)4-(3-chloro-phenylamino)-3-(4-methylsulphonylaminomethyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

n)4-(3-chloro-phenylamino)-3-(3-methylsulphonylaminomethyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

o)4-(3-chloro-phenylamino)-3-(4-formylaminomethyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

p)4-(3-chloro-phenylamino)-3-(3-formylaminomethyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

q)3-(4-acetylaminomethyl-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

r)3-(3-acetylaminomethyl-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

s)4-(3-chloro-phenylamino)-3-(4-methylsulphonylamino-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine;m.p. >260° C., APCI-MS: (M+H)⁺ =430

t)4-(3-chloro-phenylamino)-3-(3-methylsulphonylamino-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine

EXAMPLE 65

3-14-(N-BOC-Amino)-phenylaminol-4-(3-chloro-phenylamino)-1H-pyrazolo-[3,4-d]pyrimidine

With stirring, a mixture of 2 g (4.43 mmol) of1,5-dihydro-3-[4-(N-BOC-amino)-phenylamino]-4-imino-5-(3-chloro-phenyl)-4H-pyrazolo[3,4-d]pyrimidine,40 ml of dioxane and 40 ml of water is heated at 120° C. for 22 hours.Cooling to 20° C., filtering and washing the filter residue with dioxaneyield the title compound; m.p. 256-258° C.

The starting material is prepared as follows:

Step 65.1:

3-[4-(N-BOC-Amino)-phenylamino]-2-cyano-3-methylmercapto-acrylonitrile

A mixture of 13.89 g (81.58 mmol) of3,3-bis-methylmercapto-2-cyano-acrylonitrile, 17 g (81.63 mmol) ofN-BOC-1,4-phenylenediamine (Fluka) and 200 ml of methanol is heatedunder reflux for 5 hours. Cooling to room temperature, filtering andwashing the filter residue with methanol yield the title compound; m.p.190° C. Concentration of the filtrate by evaporation andrecrystallization of the residue from 50 ml of methanol yield a furtherbatch of the title compound; m.p. 190-191° C.

Step 65.2:

5-Amino-3-[4-(N-BOC-amino)-phenylamino]-4-cyano-pyrazole

Analogously to Example 49.1, the title compound [m.p. 166-168° C.] isobtained from 23.1 g (69.91 mmol) of3-[4-(N-BOC-amino)-phenylamino]-2-cyano-3-methylmercapto-acrylonitrile,4.15 ml (83.75 mmol) of hydrazine hydrate and 200 ml of methanol afterheating under reflux for 5 hours.

Step 65.3:

3-[4-(N-BOC-Amino)-phenylamino]-4-cyano-5-(ethoxy-methyleneamino)-pyrazole

With stirring, a mixture of 13.8 g (43.9 mmol) of5-amino-3-[4-(N-BOC-amino)-phenylamino]-4-cyano-pyrazole and 138 ml oforthoformic acid triethyl ester is heated at 120° C. for 3 hours, carebeing taken to ensure that the ethanol formed in the course of thereaction is distilled off from the reaction mixture. Cooling to RT,filtering and washing the filter residue with ethanol yield the titlecompound; m.p. 180-182° C.

Step 65.4:

1,5-Dihydro-3-[4-(N-BOC-amino)-phenylamino]-4-imino-5-(3-chloro-phenyl)-4H-pyrazolo[3,4-d]pyrimidine

A mixture of 7 g (18.9 mmol) of3-[4-(N-BOC-amino)-phenylamino]-4-cyano-5-(ethoxy-methyleneamino)-pyrazole,3.97 ml (37.78 mmol) of 3-chloro-aniline and 150 ml of ethanol is heatedunder reflux for 9 hours and stirred at room temperature for a further15 hours. Filtering and washing the filter residue with cold ethanolyield the title compound; m.p. 229-234° C. (decomp.); FAB-MS: (M+H)⁺=452.

EXAMPLE 66

Dry-Filled Capsules

5000 capsules, each comprising as active ingredient 0.25 g of one of thecompounds of formula I mentioned in Examples 1 to 65, are prepared asfollows:

    ______________________________________                                        Composition                                                                   ______________________________________                                        active ingredient                                                                              1250 g                                                       talcum           180 g                                                        wheat starch     120 g                                                        magnesium stearate                                                                              80 g                                                        lactose           20 g                                                        ______________________________________                                    

Preparation process: The mentioned substances are pulverized and forcedthrough a sieve of 0.6 mm mesh size. 0.33 g portions of the mixture areintroduced into gelatin capsules using a capsule-filling machine.

EXAMPLE 67

Soft Capsules

5000 soft gelatin capsules, each comprising as active ingredient 0.05 gof one of the compounds of formula I mentioned in Examples 1 to 65, areprepared as follows:

    ______________________________________                                        Composition                                                                   ______________________________________                                        active ingredient     250 g                                                   Lauroglycol            2 liters                                               ______________________________________                                    

Preparation process: The pulverized active ingredient is suspended inLauroglycol (propylene glycol laurate, Gattefosse S. A., Saint Priest,France) and ground in a wet pulveriser to a particle size of approx.from 1 to 3 mm. 0.419 g portions of the mixture are then introduced intosoft gelatin capsules using a capsule-filling machine.

EXAMPLE 68

Soft Capsules

5000 soft gelatin capsules, each comprising as active ingredient 0.05 gof one of the compounds of formula I mentioned in Examples 1 to 65, areprepared as follows:

    ______________________________________                                        Composition                                                                   ______________________________________                                               active ingredient                                                                             250 g                                                         PEG 400          1 liter                                                      Tween 80         1 liter                                               ______________________________________                                    

Preparation process: The pulverized active ingredient is suspended inPEG 400 (polyethylene glycol having an M_(r) of from approx. 380 toapprox. 420, Fluka, Switzerland) and Tween 80 (polyoxyethylene sorbitanmonolaurate, Atlas Chem. Ind. Inc., USA, supplied by Fluka, Switzerland)and ground in a wet pulverized to a particle size of approx. from 1 to 3mm. 0.43 g portions of the mixture are then introduced into soft gelatincapsules using a capsule-filling machine.

What is claimed is:
 1. A 4-amino-1H-pyrazolo[3,4-d]pyrimidine derivativeof formula I ##STR33## wherein m and n are each independently of theother an integer from 0 up to and including 3,v is 0 or 1, R is hydrogenor lower alkyl, R₁ is halogen, cyano, trifluoromethyl, hydroxy, loweralkoxy, lower alkanoyloxy, carboxy, lower alkoxycarbonyl, carbamoyl,N-lower alkylcarbamoyl, or lower alkyl that is unsubstituted orsubstituted by amino or by cyano, it being possible when several phenylsubstituents R₁ are present for those substituents to be identical ordifferent from one another, X is the group NH(CH--R₇)_(t) wherein t isan integer from 0 up to and including 3 and R₇ is hydrogen or loweralkyl, or the group (C[R₃ ]--R₄)_(q) wherein q is an integer from 0 upto and including 3 with the proviso that q and n are not simultaneously0, R₃ is hydrogen or lower alkyl and R₄ is hydrogen or lower alkyl, andR₂ is halogen, nitro, cyano, trifluoromethyl, carboxy, loweralkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, N,N-di-loweralkylcarbamoyl, azido, amino, lower alkylamino, di-lower alkylamino,di-lower alkylamino-lower alkyleneamino, benzylamino, benzoylamino,lower alkanoylamino, lower alkoxycarbonylamino, benzyloxycarbonylamino,thien-2-ylcarbonylamino, fur-2-ylcarbonylamino, pyrid-2-ylcarbonylamino,lower alkylsulfonylamino, benzenesulfonylamino, p-toluenesulfonylamino,hydroxy, lower alkanoyloxy, oxa-lower alkoxy, lower alkoxy that isunsubstituted or substituted by carboxy, lower alkoxycarbonyl, carbamoylor by N-lower alkylcarbamoyl, or lower alkyl that is unsubstituted orsubstituted by amino, lower alkanoylamino, benzoylamino, loweralkoxycarbonylamino, lower alkylsulfonylamino, benzenesulfonylamino,p-toluenesulfonylamino, cyano, hydroxy, lower alkanoyloxy, loweralkoxycarbonyloxy or by lower alkoxy, it being possible when severalphenyl substituents R₂ are present for those substituents to beidentical or different from one another and for two vicinal radicals R₂together also to form methylenedioxy, or a salt or a tautomer of such acompound.
 2. A compound of formula I according to claim 1, whereinm is 0or 1, n is an integer from 1 up to and including 3, v is 0 or 1, R ishydrogen or lower alkyl, R₁ is halogen or lower alkyl that isunsubstituted or substituted by amino or by cyano, X is the groupNH(CH--R₇)_(t) wherein t is 0 or 1 and R₇ is hydrogen or lower alkyl, orthe group (C[R₃ ]--R₄)_(q) wherein q is 0, and R₂ is halogen, nitro,carboxy, lower alkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl,N,N-di-lower alkylcarbamoyl, amino, lower alkylamino, di-loweralkylamino, di-lower alkylaminomethyleneamino, benzylamino,benzoylamino, lower alkanoylamino, lower alkoxycarbonylamino,benzyloxycarbonylamino, thien-2-ylcarbonylamino, fur-2-ylcarbonylamino,pyrid-2-ylcarbonylamino, lower alkylsulfonylamino, benzenesulfonylamino,p-toluenesulfonylamino, hydroxy, lower alkanoyloxy, lower alkoxy, orlower alkyl that is unsubstituted or substituted by amino, loweralkanoylamino, benzoylamino or by lower alkoxycarbonylamino, it beingpossible when several phenyl substituents R₂ are present for thosesubstituents to be identical or different from one another and for twovicinal radicals R₂ together also to form methylenedioxy,or a salt ofsuch a compound.
 3. A compound of formula I according to claim 1,selectedfrom3-benzylamino-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-methoxycarbonylbenzylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-carboxy-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-[3-(methylaminocarbonyl)-benzylamino]-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-[4-(methylaminocarbonyl)-benzylamino]-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(3,5-dimethoxy-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-[(3-methoxy-4-hydroxy-benzyl)-amino]-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(3-methoxycarbonyl-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3,4,5-trimethoxy-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3,4-dimethoxy-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(2,3,4-trimethoxy-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-hydroxy-4-methoxy-benzylamino)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(4-hydroxy-3,5-dimethoxy-benzylamino)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(3,4-methylenedioxy-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(2,3-methylenedioxy-benzylamino)-1H-pyrazolo[3,4-d]-pyrimidine,3-(3-chloro-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(3-chloro-4-hydroxy-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(3-chloro-4-methoxy-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-{[1-(3-chloro-phenyl)-ethyl]-amino}-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-[(1-phenyl-ethyl)-amino]-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-[3-(dimethylaminocarbonyl)-benzylamino]-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(4-methoxy-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-amino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-tert-butoxycarbonylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(3-nitro-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,3-(3-amino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-[3-methyl-butanoyl-amino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(3-[3-methyl-butanoyl-amino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(4-propanoylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-propanoylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-pivaloylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-pivaloylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-acetylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(3-acetylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-{[N,N-dimethylamino}-methyleneamino]-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-{[N,N-dimethylamino}-methyleneamino]-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-[thien-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(3-[thien-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(4-[fur-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(3-[fur-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-[pyrid-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(3-[pyrid-2-yl-carbonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(4-methylsulfonylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-methylsulfonylamino-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-[4-methyl-benzenesulfonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(3-[4-methyl-benzenesulfonylamino]-phenyl)-1H-pyrazolo[3,4-d]-pyrimidine,3-(3-tert-butoxycarbonylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,3-(4-benzyloxycarbonylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,3-(3-benzyloxycarbonylamino-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,3-(4-acetylamino-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-dimethylamino-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-methoxy-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-methoxy-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-benzylamino-3-(3-methyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,(S)-3-(3-methyl-phenylamino)-4-(1-phenyl-ethylamino)-1H-pyrazolo[3,4-d]pyrimidine,(R)-3-(3-methyl-phenylamino)-4-(1-phenyl-ethylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-benzylamino-3-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-methoxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(3-hydroxy-phenyl)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-hydroxy-phenyl)-1H-pyrazolo[3,4-d]pyrimidine,(R)-3-(3-chloro-phenylamino)-4-(1-phenyl-ethylamino)-1H-pyrazolo[3,4-d]pyrimidine,(S)-3-(3-chloro-phenylamino)-4-(1-phenyl-ethylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-acetylamino-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-amino-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-hydroxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-amino-benzylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-ethoxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3,4-dimethoxy-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3,5-dimethoxy-phenylamino)-1H-pyrazolo[3,4-d]pynimidine,4-(3-chloro-phenylamino)-3-(3-formylamino-4-methoxy-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,3-(3-acetylamino-4-methoxy-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,3-(4-acetylamino-3-methoxy-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,4-(3-chloro-phenylamino)-3-(4-formylamino-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-formylamino-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-propionylamino-benzylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-aminomethyl-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(3-aminomethyl-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-[4-(N-BOC-aminomethyl)-phenylamino]-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,3-[3-(N-BOC-aminomethyl)-phenylamino]-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]-pyrimidine,3-[4-(N-BOC-amino)-phenylamino]-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-methylsulphonylaminomethyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-methylsulphonylaminomethyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(4-formylaminomethyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-chloro-phenylamino)-3-(3-formylaminomethyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-acetylaminomethyl-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,and3-(3-acetylaminomethyl-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidineora pharmaceutically acceptable salt.
 4. A4-phenylamino-1H-pyrazolo[3,4-d]pyrimidine derivative of formula Iaaccording to claim 1 ##STR34## wherein m and n are each independently ofthe other an integer from 0 up to and including 3,R₁ is halogen, cyano,trifluoromethyl, hydroxy, lower alkoxy, lower alkanoyloxy, carboxy,lower alkoxycarbonyl, carbamoyl, N-lower alkylcarbamoyl, or lower alkylthat is unsubstituted or substituted by amino or by cyano, it beingpossible when several phenyl substituents R₁ are present for thosesubstituents to be identical or different from one another, X is thegroup NH(CH₂)_(t) wherein t is an integer from 0 up to and including 3,or the group (C[R₃ ]--R₄)_(q) wherein q is an integer from 0 up to andincluding 3 with the proviso that q and n are not simultaneously 0, R₃is hydrogen or lower alkyl and R₄ is hydrogen or lower alkyl, and R₂ ishalogen, nitro, cyano, trifluoromethyl, carboxy, lower alkoxycarbonyl,carbamoyl, N-lower alkylcarbamoyl, amino, lower alkylamino, hydroxy,lower alkanoyloxy, oxa-lower alkoxy, lower alkoxy that is unsubstitutedor substituted by carboxy, lower alkoxycarbonyl, carbamoyl or by N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by hydroxy, it being possible when several phenyl substituentsR₂ are present for those substituents to be identical or different fromone another,or a salt of such a compound.
 5. A compound of formula Iaaccording to claim 4, wherein X is the group NH(CH₂)_(t) wherein t is aninteger from 0 up to and including 3, or the group (C[R₃ ]--R₄)_(q)wherein q is 0, or a salt of such a compound.
 6. A compound of formulaIa according to claim 4, wherein X is the group NH(CH₂)_(t) wherein t is0, or a salt of such a compound.
 7. A compound of formula Ia accordingto claim 4, wherein m and n are each independently of the other 0 or 1,R₁ is halogen, or lower alkyl that is unsubstituted or substituted byamino or by cyano, X is the group NH(CH₂)_(t) wherein t is 0, and R₂ ishalogen or lower alkoxy, or a salt of such a compound.
 8. A compound offormula Ia according to claim 4, selectedfrom3,4-diphenylamino-1H-pyrazolo[3,4-d]pyrimidine,3-(3-chloro-phenylamino)-4-phenylamino-1H-pyrazolo[3,4-d]pyrimidine,3,4-di(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-bromo-phenylamino)-3-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(3-chloro-phenylamino)-4-(3-methyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-[2-cyano-ethyl]-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidine,4-(4-[2-cyano-ethyl]-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidine,4-(3-cyanomethyl-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidine,3-(3-chloro-phenylamino)-4-(3-methyl-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,4-(3-[3-amino-propyl]-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidine,4-(4-[3-amino-propyl]-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidine,4-(3-[2-amino-ethyl]-phenylamino)-3-phenylamino-1H-pyrazolo[3,4-d]pyrimidine,3-(4-chloro-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(3-fluoro-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-fluoro-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-methoxy-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidine,3-(4-hydroxy-phenylamino)-4-(3-chloro-phenylamino)-1H-pyrazolo[3,4-d]pyrimidineora pharmaceutically acceptable salt.
 9. A pharmaceutical compositioncomprising a pharmaceutically acceptable carrier or diluent and atherapeutically effective amount of a compound of claim 1, or apharmaceutically acceptable salt thereof.
 10. A method of inhibitingprotein tyrosine kinases in a warm-blooded animal comprisingadministering to said warm-blooded animal a therapeutically effectiveamount of a 4-amino-1H-pyrazolo[3,4-d]pyrimidine derivative of formula I##STR35## wherein m and n are each independently of the other an integerfrom 0 up to and including 3,v is 0 or 1, R is hydrogen or lower alkyl,R₁ is halogen, cyano, trifluoromethyl, hydroxy, lower alkoxy, loweralkanoyloxy, carboxy, lower alkoxycarbonyl, carbamoyl, N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by cyano, it being possible when several phenyl substituents R₁are present for those substituents to be identical or different from oneanother, X is the group NH(CH--R₇)_(t) wherein t is an integer from 0 upto and including 3 and R₇ is hydrogen or lower alkyl, or the group (C[R₃]--R₄)_(q) wherein q is an integer from 0 up to and including 3, R₃ ishydrogen or lower alkyl and R₄ is hydrogen or lower alkyl, and R₂ ishalogen, nitro, cyano, trifluoromethyl, carboxy, lower alkoxycarbonyl,carbamoyl, N-lower alkylcarbamoyl, N,N-di-lower alkylcarbamoyl, azido,amino, lower alkylamino, di-lower alkylamino, di-lower alkylamino-loweralkyleneamino, benzylamino, benzoylamino, lower alkanoylamino, loweralkoxycarbonylamino, benzyloxycarbonylamino, thien-2-ylcarbonylamino,fur-2-ylcarbonylamino, pyrid-2-ylcarbonylamino, loweralkylsulfonylamino, benzenesulfonylamino, p-toluenesulfonylamino,hydroxy, lower alkanoyloxy, oxa-lower alkoxy, lower alkoxy that isunsubstituted or substituted by carboxy, lower alkoxycarbonyl, carbamoylor by N-lower alkylcarbamoyl, or lower alkyl that is unsubstituted orsubstituted by amino, lower alkanoylamino, benzoylamino, loweralkoxycarbonylamino, lower alkylsulfonylamino, benzenesulfonylamino,p-toluenesulfonylamino, cyano, hydroxy, lower alkanoyloxy, loweralkoxycarbonyloxy or by lower alkoxy, it being possible when severalphenyl substituents R₂ are present for those substituents to beidentical or different from one another and for two vicinal radicals R₂together also to form methylenedioxy, or a pharmaceutically acceptablesalt thereof.
 11. A method of treating tumors mediated by proteintyrosine kinase inhibition in a warm-blooded animal comprisingadministering to said warm-blooded animal a therapeutically effectiveamount of a 4-amino-1H-pyrazolo[3,4-d]pyrimidine derivative of formula I##STR36## wherein m and n are each independently of the other an integerfrom 0 up to and including 3,v is 0 or 1, R is hydrogen or lower alkyl,R₁ is halogen, cyano, trifluoromethyl, hydroxy, lower alkoxy, loweralkanoyloxy, carboxy, lower alkoxycarbonyl, carbamoyl, N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by cyano, it being possible when several phenyl substituents R₁are present for those substituents to be identical or different from oneanother, X is the group NH(CH--R₇)_(t) wherein t is an integer from 0 upto and including 3 and R₇ is hydrogen or lower alkyl, or the group (C[R₃]--R₄)_(q) wherein q is an integer from 0 up to and including 3, R₃ ishydrogen or lower alkyl and R₄ is hydrogen or lower alkyl, and R₂ ishalogen, nitro, cyano, trifluoromethyl, carboxy, lower alkoxycarbonyl,carbamoyl, N-lower alkylcarbamoyl, N,N-di-lower alkylcarbamoyl, azido,amino, lower alkylamino, di-lower alkylamino, di-lower alkylamino-loweralkyleneamino, benzylamino, benzoylamino, lower alkanoylamino, loweralkoxycarbonylamino, benzyloxycarbonylamino, thien-2-ylcarbonylamino,fur-2-ylcarbonylamino, pyrid-2-ylcarbonylamino, loweralkylsulfonylamino, benzenesulfonylamino, p-toluenesulfonylamino,hydroxy, lower alkanoyloxy, oxa-lower alkoxy, lower alkoxy that isunsubstituted or substituted by carboxy, lower alkoxycarbonyl, carbamoylor by N-lower alkylcarbamoyl, or lower alkyl that is unsubstituted orsubstituted by amino, lower alkanoylamino, benzoylamino, loweralkoxycarbonylamino, lower alkylsulfonylamino, benzenesulfonylamino,p-toluenesulfonylamino, cyano, hydroxy, lower alkanoyloxy, loweralkoxycarbonyloxy or by lower alkoxy, it being possible when severalphenyl substituents R₂ are present for those substituents to beidentical or different from one another and for two vicinal radicals R₂together also to form methylenedioxy, or a pharmaceutically acceptablesalt thereof.
 12. A process for preparing a4-amino-1H-pyrazolo[3,4-d]pyrimidine derivative of formula I ##STR37##wherein m and n are each independently of the other an integer from 0 upto and including 3,v is 0 or 1, R is hydrogen or lower alkyl, R₁ ishalogen, cyano, trifluoromethyl, hydroxy, lower alkoxy, loweralkanoyloxy, carboxy, lower alkoxycarbonyl, carbamoyl, N-loweralkylcarbamoyl, or lower alkyl that is unsubstituted or substituted byamino or by cyano, it being possible when several phenyl substituents R₁are present for those substituents to be identical or different from oneanother, X is the group NH(CH--R₇)_(t) wherein t is an integer from 0 upto and including 3 and R₇ is hydrogen or lower alkyl, or the group (C[R₃]--R₄)_(q) wherein q is an integer from 0 up to and including 3, R₃ ishydrogen or lower alkyl and R₄ is hydrogen or lower alkyl, and R₂ ishalogen, nitro, cyano, trifluoromethyl, carboxy, lower alkoxycarbonyl,carbamoyl, N-lower alkylcarbamoyl, N,N-di-lower alkylcarbamoyl, azido,amino, lower alkylamino, di-lower alkylamino, di-lower alkylamino-loweralkyleneamino, benzylamino, benzoylamino, lower alkanoylamino, loweralkoxycarbonylamino, benzyloxycarbonylamino, thien-2-ylcarbonylamino,fur-2-ylcarbonylamino, pyrid-2-ylcarbonylamino, loweralkylsulfonylamino, benzenesulfonylamino, p-toluenesulfonylamino,hydroxy, lower alkanoyloxy, oxa-lower alkoxy, lower alkoxy that isunsubstituted or substituted by carboxy, lower alkoxycarbonyl, carbamoylor by N-lower alkylcarbamoyl, or lower alkyl that is unsubstituted orsubstituted by amino, lower alkanoylamino, benzoylamino, loweralkoxycarbonylamino, lower alkylsulfonylamino, benzenesulfonylamino,p-toluenesulfonylamino, cyano, hydroxy, lower alkanoyloxy, loweralkoxycarbonyloxy or by lower alkoxy, it being possible when severalphenyl substituents R₂ are present for those substituents to beidentical or different from one another and for two vicinal radicals R₂together also to form methylenedioxy, or a salt of such a compound,which process comprises treating a compound of formula II ##STR38##wherein R₅ is hydrogen or methyl, R₆ is alkoxy having from 1 to 3 carbonatoms or is nitro, r is an integer from 0 to 2, and the remainingsubstituents and symbols are as defined above, with a suitable Lewisacid, and, if desired, converting a compound of formula I into its salt,or converting an obtainable salt of a compound of formula I into thefree compound.